Devices and systems for receiving biological material and providing enriched components thereof

Abstract

This document provides devices and systems (e.g., point of care devices and systems) designed to receive biological material (e.g., fat tissue) from a mammal (e.g., a human or a horse) and designed to process that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal. For example, point of care devices designed to harvest fat tissue from a mammal (e.g., a human or a horse) in a clinical setting and to process that fat tissue to provide adipose-derived stem cells as an enriched end product of the harvested fat tissue that can, optionally, be reintroduced into that same mammal in the clinical setting are provided. Methods for using the devices and systems described herein also are provided.

Description

[0001] DEVICES AND SYSTEMS FOR RECEIVING BIOLOGICAL MATERIAL AND PROVIDING ENRICHED COMPONENTS THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority from U.S. Provisional Application Serial No. 63 / 527,149, filed July 17, 2023. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application.

[0004] TECHNICAL FIELD

[0005] This document relates to devices and systems (e.g., point of care devices and systems) designed to receive biological material (e.g., fat tissue) from a mammal (e.g., a human or a horse) and designed to process that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal. For example, this document provides point of care devices designed to harvest fat tissue from a mammal (e.g., a human) in a clinical setting and to process that fat tissue to provide adipose- derived stem cells as an enriched end product of the harvested fat tissue that can, optionally, be reintroduced into that same mammal in the clinical setting. This document also provides methods for obtaining biological material (e.g., fat tissue) from a mammal (e.g., a human) and processing that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal.

[0006] BACKGROUND

[0007] Adipose-derived stem cells (also referred to as adipose-derived regenerative cells, adipose-derived adult stem cells, and adipose mesenchymal stem cells) and stromal vascular fractions both have raised interest in therapeutic applications in the areas of regenerative medicine and tissue engineering. Studies described elsewhere (Mizuno et al., Stem Cells, 30(5):804-810, 2012; and Si et al., Biomedicine & Pharmacotherapy, 114: 108765, 2019) demonstrated the potential of adipose-derived stem cells in regenerative medicine in general and specifically in, for example, bone / chondral repair, muscle regeneration, treatment of osteoarthritis (Lee t al., Stem Cells Translational Medicine, 8(6):504-511, 2019; and Song et al., Regenerative Medicine, 13(3):295-307, 2018), treatment of rheumatic diseases, chronic wound healing, treatment of diabetic foot (Gadelkarim et al., Biomedicine & Pharmacotherapy, 107:625-633, 2018), hair growth (Tak et al., Stem Cells Translational Medicine, 9(8):839-849, 2020), treatment of multiple sclerosis (Ganji et al., Journal of Molecular Neuroscience : MN, 70(7): 1088- 1099, 2020), and the treatment of many more conditions. The interest in these multipotent somatic stem cells and the stromal vascular fractions is based on the possibility of obtaining many cells and materials by means of minimally invasive procedures. Other forms of stem cell therapy such as bone marrow stem cell therapies and umbilical stem cell therapies can have higher risks associated with harvesting, can have considerable morbidity, can have difficulties obtaining significant quantities of cells, and can raise difficult ethical and regulatory questions. On the other hand, adipose-derived stem cells can be obtained not only from solid fat but also from lipoaspirate. The absence of an immune response, the ability of the cells to self-renew, their resemblance to a natural healing process, and the ability of the cells to develop into a desired tissue is of particular interest. The ability of adipose-derived stem cells to differentiate into different tissuespecific progenitors, to migrate into damaged sites, and to act through autocrine and paracrine pathways demonstrate that these cells could fulfill the criteria for a successful cell therapy.

[0008] SUMMARY

[0009] The absence of standardization protocols in cell management within laboratories and surgical suites, as well as the non-sterile, complicated, and slow process of adipose- derived stem cell isolation, has limited their widespread use by balancing their real-life benefit versus patient safety and security.

[0010] This document provides devices and systems (e.g., point of care devices and systems) designed to receive biological material (e.g., fat tissue) from a mammal (e.g., a human) and designed to process that biological material to provide an end product that includes one or more enriched components (e.g., stem cells) of the received biological material that can, optionally, be reintroduced into that same mammal. For example, this document provides point of care devices designed to harvest fat tissue from a mammal (e.g., a human) in a clinical setting and to process that fat tissue to provide adipose- derived stem cells as an enriched end product of the harvested fat tissue that can, optionally, be reintroduced into that same mammal in the clinical setting. This document also provides methods for obtaining biological material (e.g., fat tissue) from a mammal (e.g., a human) and processing that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal.

[0011] As described herein, a harvesting system can be designed to harvest biological material (e.g., fat tissue) from a mammal (e.g., a human) and process that biological material to form a viscous fluid that includes intact cells. The system also can be designed to process that viscous fluid to form an end product enriched for one or more components of the harvested biological material, such as adipose-derived stem cells. In some cases, the end product can be directly administered back into the same mammal from which the biological material was harvested. For example, in a clinical setting, a closed loop system provided herein can be used to harvest biological material (e.g., fat tissue) from a human, to process that harvested biological material to obtain an end product of enriched adipose-derived stem cells, and to administer that obtained end product directly back into that human.

[0012] In general, one aspect of this document features a device for obtaining an enriched population of cells from biological material obtained from a mammal. The device comprises (or consists essentially of, or consists of) (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the enriched population from the conduit, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of cells from the dispersed form of the biological material, wherein the enriched population of cells comprises a greater number of cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material. The enriched population of cells can be an enriched population of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells can be adipose-derived stem cells. The biological material can comprise fat tissue. The mammal can be a human. The mammal can be a horse. The conduit can comprise flexible tubing. The conduit can be from about 20 cm to about 10 m in length. The conduit can comprise at least one section having an inner diameter of about 1 mm to 5 mm. In some cases, a maximum inner diameter along any point of the conduit can be no greater than 20 mm. In some cases, a minimum inner diameter along any point of the conduit can be no less than 0.1 mm. The conduit can comprise stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof. The inlet port can be configured to attach to an outlet of a liposuction device. The device can comprise an adaptor configured to attach the inlet port to the liposuction device. The inlet port can be configured to collect the biological material directly from the mammal. The outlet port can be configured to attach to an inlet of an administering device. The outlet port can be configured to deliver the enriched population directly to the mammal. At least one of the one or more pumping elements can be a displacement pump. The displacement pump can be a reciprocating pump. The reciprocating pump can be a diaphragm pump. The displacement pump can be a rotary or continuous pump. The displacement pump can be a peristaltic pump. In some cases, no component of any of the one or more pumping elements can be located within the conduit. In some cases, no component of any of the one or more pumping elements can contact the biological material or the dispersed form of the biological material when the biological material and the dispersed form of the biological material are within the conduit. The one or more dispersing elements can be bottlenecks in the conduit. The bottlenecks can be compressed sections of the conduit. The conduit can comprise two or more bottlenecks separated by intervening segments of the conduit, and wherein the bottlenecks can have a reduced maximum diameter as compared to the intervening segments. The conduit can comprise 5 to 40 bottlenecks. The conduit can comprise 20 to 30 bottlenecks. The conduit can be configured as a series of loops. Each loop can comprise 4 to 6 bottlenecks. Each loop can be configured to interact with a pumping element. At least one of the one or more dispersing elements can comprise one or more microstructures on an inner surface of the conduit. At least one of one or more separating elements can be a filter. The filter can be a tangential flow filter. The tangential flow filter can be a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores. The one or more separating elements can be filters in an axial arrangement. The one or more separating elements can be filters in a radial arrangement. The one or more separating elements can be filters in a parallel arrangement. The one or more separating elements can be filters in a serial arrangement. The one or more separating elements can be filters in a serial and parallel arrangement. The one or more separating elements can comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger than about 80 to 100 pm if present in the dispersed form of the biological material from the enriched population. The larger items can comprise adipose cells. The one or more separating elements can comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in the dispersed form of the biological material from the enriched population. The smaller items can comprise fat globules, erythrocytes, or fat globules and erythrocytes. The device can comprise one or more first holding chambers located along the conduit between the inlet port and a first of the one or more dispersing elements and configured to accumulate and hold the biological material. The device can comprise one or more second holding chambers located along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to accumulate and hold the dispersed form of the biological material. The device can comprise one or more third holding chambers located along the conduit between a first of the one or more separating elements and the outlet port and configured to accumulate and hold the enriched population. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between the inlet port and a first of the one or more dispersing elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more separating elements and the outlet port and configured to add a fluid to the enriched population. The fluid can be water or saline. The device can lack any inflow ports that fluidly connect to the conduit along the conduit between the inlet port and the outlet port. The device can comprise no more than one inflow port fluidly connected to the conduit along the conduit between the inlet port and the outlet port. The enriched population can comprise about 150,000 cells / cm3. The enriched population of cells can comprise at least 5-fold to 10-fold more cells per mg of starting biological material than the dispersed form of the biological material.

[0013] In another aspect, this document features a device for obtaining an enriched population of adipose-derived stem cells from biological material obtained from a mammal and returning the enriched population to the mammal. The device comprises (or consists essentially of, or consists of) (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to deliver the enriched population to the mammal, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact adipose-derived stem cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of adipose-derived stem cells from the dispersed form of the biological material, wherein the enriched population of adipose-derived stem cells comprises a greater number of adipose-derived stem cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material. The enriched population of cells can be an enriched population of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells can be adipose-derived stem cells. The biological material can comprise fat tissue. The mammal can be a human. The conduit can comprise flexible tubing. The conduit can be from about 20 cm to about 10 m in length. The conduit can comprise at least one section having an inner diameter of about 1 mm to 5 mm. In some cases, a maximum inner diameter along any point of the conduit can be no greater than 20 mm. In some cases, a minimum inner diameter along any point of the conduit can be no less than 0.1 mm. The conduit can comprise stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof. The inlet port can be configured to attach to an outlet of a liposuction device. The device can comprise an adaptor configured to attach the inlet port to the liposuction device. The inlet port can be configured to collect the biological material directly from the mammal. The outlet port can be configured to attach to an inlet of an administering device. The outlet port can be configured to deliver the enriched population directly to the mammal. At least one of the one or more pumping elements can be a displacement pump. The displacement pump can be a reciprocating pump. The reciprocating pump can be a diaphragm pump. The displacement pump can be a rotary or continuous pump. The displacement pump can be a peristaltic pump. In some cases, no component of any of the one or more pumping elements can be located within the conduit. In some cases, no component of any of the one or more pumping elements can contact the biological material or the dispersed form of the biological material when the biological material and the dispersed form of the biological material are within the conduit. The one or more dispersing elements can be bottlenecks in the conduit. The bottlenecks can be compressed sections of the conduit. The conduit can comprise two or more bottlenecks separated by intervening segments of the conduit, and wherein the bottlenecks can have a reduced maximum diameter as compared to the intervening segments. The conduit can comprise 5 to 40 bottlenecks. The conduit can comprise 20 to 30 bottlenecks. The conduit can be configured as a series of loops. Each loop can comprise 4 to 6 bottlenecks. Each loop can be configured to interact with a pumping element. At least one of the one or more dispersing elements can comprise one or more microstructures on an inner surface of the conduit. At least one of one or more separating elements can be a filter. The filter can be a tangential flow filter. The tangential flow filter can be a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores. The one or more separating elements can be filters in an axial arrangement. The one or more separating elements can be filters in a radial arrangement. The one or more separating elements can be filters in a parallel arrangement. The one or more separating elements can be filters in a serial arrangement. The one or more separating elements can be filters in a serial and parallel arrangement. The one or more separating elements can comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger than about 80 to 100 pm if present in the dispersed form of the biological material from the enriched population. The larger items can comprise adipose cells. The one or more separating elements can comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in the dispersed form of the biological material from the enriched population. The smaller items can comprise fat globules, erythrocytes, or fat globules and erythrocytes. The device can comprise one or more first holding chambers located along the conduit between the inlet port and a first of the one or more dispersing elements and configured to accumulate and hold the biological material. The device can comprise one or more second holding chambers located along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to accumulate and hold the dispersed form of the biological material. The device can comprise one or more third holding chambers located along the conduit between a first of the one or more separating elements and the outlet port and configured to accumulate and hold the enriched population. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between the inlet port and a first of the one or more dispersing elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more separating elements and the outlet port and configured to add a fluid to the enriched population. The fluid can be water or saline. The device can lack any inflow ports that fluidly connect to the conduit along the conduit between the inlet port and the outlet port. The device can comprise no more than one inflow port fluidly connected to the conduit along the conduit between the inlet port and the outlet port. The enriched population can comprise about 150,000 cells / cm3. The enriched population of cells can comprise at least 5-fold to 10-fold more cells per mg of starting biological material than the dispersed form of the biological material.

[0014] In another aspect, this document features a method for preparing an enriched population of cells using a device for obtaining an enriched population of cells from biological material obtained from a mammal. The device can comprise (or can consist essentially of, or can consist of) (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the enriched population from the conduit, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of cells from the dispersed form of the biological material, wherein the enriched population of cells comprises a greater number of cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material. The enriched population of cells can be an enriched population of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells can be adipose-derived stem cells. The biological material can comprise fat tissue. The mammal can be a human. The conduit can comprise flexible tubing. The conduit can be from about 20 cm to about 10 m in length. The conduit can comprise at least one section having an inner diameter of about 1 mm to 5 mm. In some cases, a maximum inner diameter along any point of the conduit can be no greater than 20 mm. In some cases, a minimum inner diameter along any point of the conduit can be no less than 0.1 mm. The conduit can comprise stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof. The inlet port can be configured to attach to an outlet of a liposuction device. The device can comprise an adaptor configured to attach the inlet port to the liposuction device. The inlet port can be configured to collect the biological material directly from the mammal. The outlet port can be configured to attach to an inlet of an administering device. The outlet port can be configured to deliver the enriched population directly to the mammal. At least one of the one or more pumping elements can be a displacement pump. The displacement pump can be a reciprocating pump. The reciprocating pump can be a diaphragm pump. The displacement pump can be a rotary or continuous pump. The displacement pump can be a peristaltic pump. In some cases, no component of any of the one or more pumping elements can be located within the conduit. In some cases, no component of any of the one or more pumping elements can contact the biological material or the dispersed form of the biological material when the biological material and the dispersed form of the biological material are within the conduit. The one or more dispersing elements can be bottlenecks in the conduit. The bottlenecks can be compressed sections of the conduit. The conduit can comprise two or more bottlenecks separated by intervening segments of the conduit, and wherein the bottlenecks can have a reduced maximum diameter as compared to the intervening segments. The conduit can comprise 5 to 40 bottlenecks. The conduit can comprise 20 to 30 bottlenecks. The conduit can be configured as a series of loops. Each loop can comprise 4 to 6 bottlenecks. Each loop can be configured to interact with a pumping element. At least one of the one or more dispersing elements can comprise one or more microstructures on an inner surface of the conduit. At least one of one or more separating elements can be a filter. The filter can be a tangential flow filter. The tangential flow filter can be a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores. The one or more separating elements can be filters in an axial arrangement. The one or more separating elements can be filters in a radial arrangement. The one or more separating elements can be filters in a parallel arrangement. The one or more separating elements can be filters in a serial arrangement. The one or more separating elements can be filters in a serial and parallel arrangement. The one or more separating elements can comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger than about 80 to 100 pm if present in the dispersed form of the biological material from the enriched population. The larger items can comprise adipose cells. The one or more separating elements can comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in the dispersed form of the biological material from the enriched population. The smaller items can comprise fat globules, erythrocytes, or fat globules and erythrocytes. The device can comprise one or more first holding chambers located along the conduit between the inlet port and a first of the one or more dispersing elements and configured to accumulate and hold the biological material. The device can comprise one or more second holding chambers located along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to accumulate and hold the dispersed form of the biological material. The device can comprise one or more third holding chambers located along the conduit between a first of the one or more separating elements and the outlet port and configured to accumulate and hold the enriched population. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between the inlet port and a first of the one or more dispersing elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more separating elements and the outlet port and configured to add a fluid to the enriched population. The fluid can be water or saline. The device can lack any inflow ports that fluidly connect to the conduit along the conduit between the inlet port and the outlet port. The device can comprise no more than one inflow port fluidly connected to the conduit along the conduit between the inlet port and the outlet port. The enriched population can comprise about 150,000 cells / cm3. The enriched population of cells can comprise at least 5-fold to 10-fold more cells per mg of starting biological material than the dispersed form of the biological material. The method comprises (or consists essentially of, or consists of): (a) passing the biological material of the mammal through the inlet port and into the conduit, (b) actuating the one or more pumping elements to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) actuating the one or more dispersing elements to form the dispersed form of the biological material from the biological material, and (d) allowing the one or more separating elements to form the enriched population of cells from the dispersed form of the biological material. The biological material can be passed through the inlet port and into the conduit without purifying the biological material, without adding any preservatives to the biological material, and without diluting the biological material. The method can comprise passing the biological material through the inlet port and into the conduit no more than about 5 minutes after the biological material was removed from the mammal. The method can be a method that does not comprise centrifuging the biological material or the enriched population of cells.

[0015] In another aspect, this document features a method for delivering cells to a mammal using a device. The device can comprise (or can consist essentially of, or can consist of) (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the enriched population from the conduit, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of cells from the dispersed form of the biological material, wherein the enriched population of cells comprises a greater number of cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material. The enriched population of cells can be an enriched population of adipose- derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells can be adipose-derived stem cells. The biological material can comprise fat tissue. The mammal can be a human. The conduit can comprise flexible tubing. The conduit can be from about 20 cm to about 10 m in length. The conduit can comprise at least one section having an inner diameter of about 1 mm to 5 mm. In some cases, a maximum inner diameter along any point of the conduit can be no greater than 20 mm. In some cases, a minimum inner diameter along any point of the conduit can be no less than 0.1 mm. The conduit can comprise stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof. The inlet port can be configured to attach to an outlet of a liposuction device. The device can comprise an adaptor configured to attach the inlet port to the liposuction device. The inlet port can be configured to collect the biological material directly from the mammal. The outlet port can be configured to attach to an inlet of an administering device. The outlet port can be configured to deliver the enriched population directly to the mammal. At least one of the one or more pumping elements can be a displacement pump. The displacement pump can be a reciprocating pump. The reciprocating pump can be a diaphragm pump. The displacement pump can be a rotary or continuous pump. The displacement pump can be a peristaltic pump. In some cases, no component of any of the one or more pumping elements can be located within the conduit. In some cases, no component of any of the one or more pumping elements can contact the biological material or the dispersed form of the biological material when the biological material and the dispersed form of the biological material are within the conduit. The one or more dispersing elements can be bottlenecks in the conduit. The bottlenecks can be compressed sections of the conduit. The conduit can comprise two or more bottlenecks separated by intervening segments of the conduit, and wherein the bottlenecks can have a reduced maximum diameter as compared to the intervening segments. The conduit can comprise 5 to 40 bottlenecks. The conduit can comprise 20 to 30 bottlenecks. The conduit can be configured as a series of loops. Each loop can comprise 4 to 6 bottlenecks. Each loop can be configured to interact with a pumping element. At least one of the one or more dispersing elements can comprise one or more microstructures on an inner surface of the conduit. At least one of one or more separating elements can be a filter. The filter can be a tangential flow filter. The tangential flow filter can be a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores. The one or more separating elements can be filters in an axial arrangement. The one or more separating elements can be filters in a radial arrangement. The one or more separating elements can be filters in a parallel arrangement. The one or more separating elements can be filters in a serial arrangement. The one or more separating elements can be filters in a serial and parallel arrangement. The one or more separating elements can comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger than about 80 to 100 pm if present in the dispersed form of the biological material from the enriched population. The larger items can comprise adipose cells. The one or more separating elements can comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in the dispersed form of the biological material from the enriched population. The smaller items can comprise fat globules, erythrocytes, or fat globules and erythrocytes. The device can comprise one or more first holding chambers located along the conduit between the inlet port and a first of the one or more dispersing elements and configured to accumulate and hold the biological material. The device can comprise one or more second holding chambers located along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to accumulate and hold the dispersed form of the biological material. The device can comprise one or more third holding chambers located along the conduit between a first of the one or more separating elements and the outlet port and configured to accumulate and hold the enriched population. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between the inlet port and a first of the one or more dispersing elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to add a fluid to the biological material. The fluid can be water or saline. The device can comprise one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more separating elements and the outlet port and configured to add a fluid to the enriched population. The fluid can be water or saline. The device can lack any inflow ports that fluidly connect to the conduit along the conduit between the inlet port and the outlet port. The device can comprise no more than one inflow port fluidly connected to the conduit along the conduit between the inlet port and the outlet port. The enriched population can comprise about 150,000 cells / cm3. The enriched population of cells can comprise at least 5-fold to 10-fold more cells per mg of starting biological material than the dispersed form of the biological material. The method comprises (or consists essentially of, or consists of) (a) passing the biological material of the mammal through the inlet port and into the conduit, (b) actuating the one or more pumping elements to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) actuating the one or more dispersing elements to form the dispersed form of the biological material from the biological material, (d) allowing the one or more separating elements to form the enriched population of cells from the dispersed form of the biological material, and (e) administering the enriched population of cells to the mammal. The biological material can be passed through the inlet port and into the conduit without purifying the biological material, without adding any preservatives to the biological material, and without diluting the biological material. The method can comprise passing the biological material through the inlet port and into the conduit no more than about 5 minutes after the biological material was removed from the mammal. The method can be a method that does not comprise centrifuging the biological material or the enriched population of cells. The enriched population of cells can be administered to the mammal without exposing the enriched population of cells to open air. The method can comprise administering the enriched population of cells to the mammal within about 5 to about 30 minutes after the biological material was obtained from the mammal. The method can be a method that does not comprise culturing the biological materials or the enriched population of cells.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0017] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0018] DESCRIPTION OF DRAWINGS

[0019] Figure l is a diagram of a device for harvesting biological material from a mammal and processing the biological material to form a sample enriched for one or more components of the harvested biological material, according to some embodiments. Figure 2 is a diagram of another device, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to form a sample enriched for one or more components of the harvested biological material.

[0020] Figure 3 is a diagram of another device, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to form a sample enriched for one or more components of the harvested biological material.

[0021] Figure 4 is a diagram of another device, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to form a sample enriched for one or more components of the harvested biological material.

[0022] Figure 5 is a diagram of another device, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to a sample enriched for one or more components of the harvested biological material.

[0023] Figures 6A and 6B show a device for harvesting biological material from a mammal and processing the biological material to enrich for one or more components of the harvested biological material, according to some embodiments.

[0024] Figure 7 is a flow chart of a method, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to enrich for one or more components of the harvested biological material.

[0025] Figure 8 is a flow chart of a method, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to enrich for one or more components of the harvested biological material.

[0026] Figure 9 is a flow chart of a method, according to some embodiments, for harvesting biological material from a mammal and processing the biological material to enrich for one or more components of the harvested biological material.

[0027] Figure 10 is a diagram depicting representative elements and functionality of a peristaltic pump, according to some embodiments.

[0028] Figure 11 is a diagram depicting an embodiment of a peristaltic pump having integrated bottlenecks generated by spring-loaded elements positioned on an exterior surface of the conduit. Figure 12 is a diagram depicting an embodiment of a peristaltic pump having integrated bottlenecks generated by radially movable elements positioned on the pump impeller.

[0029] Figure 13 is a diagram depicting an embodiment of a peristaltic pump having integrated bottlenecks generated by spring-loaded elements positioned on an exterior surface of the conduit and by radially movable roller elements positioned on the pump impeller.

[0030] Figure 14 is a diagram depicting a device provided herein, according to some embodiments, indicating the flow of biological material through the device.

[0031] Figure 15 includes a diagram showing a blown up view (indicated by the dashed circle and line in the diagram at the left) of a first filter stage in a device provided herein, according to some embodiments.

[0032] Figure 16 includes a diagram showing a blown up view (indicated by the dashed circle and line in the diagram at the left) of a first filter stage in a device provided herein, according to some embodiments.

[0033] Figure 17 includes a diagram showing a blown up view (indicated by the dashed circle and line in the diagram at the upper left) of a second filter stage in a device provided herein, according to some embodiments.

[0034] Figure 18 includes a diagram showing a blown up view (indicated by the dashed circle in the diagram at the left) of an outlet for a retentate in a device provided herein, according to some embodiments.

[0035] Like reference symbols in the various drawings indicate like elements.

[0036] DETAILED DESCRIPTION

[0037] This document provides devices and systems (e.g., point of care devices and systems) designed to receive biological material (e.g., fat tissue) from a mammal (e.g., a human) and designed to process that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal. For example, this document provides point of care devices designed to harvest fat tissue from a mammal (e.g., a human or a horse) in any appropriate setting (e.g., a clinical setting appropriate for a human or a nonhuman mammal such as, without limitation, a horse) and to process that fat tissue to provide adipose-derived stem cells as an enriched end product of the harvested fat tissue that can, optionally, be reintroduced into that same mammal in the same setting (e.g., the clinical setting). This document also provides methods for obtaining biological material (e.g., fat tissue) from a mammal (e.g., a human) and processing that biological material to provide an end product that includes one or more enriched components of the received biological material that can, optionally, be reintroduced into that same mammal. In some cases, the methods provided herein can be used to generate an enriched cell population with no centrifugation. Thus, biological material can be obtained from a mammal and can be processed to form, for example, an enriched population of cells that can be reintroduced into the same mammal without having undergone centrifugation.

[0038] In some cases, the end product containing enriched material can contain from about 1 x 103to about 1 x 108(e.g., from about 7.5 x 104to about 1 x 107, from about 1 x 105to about 2 x 105, from about 2 x 105to about 5 x 105, from about 5 x 105to about 1 x 106, from about 1 x 106to about 5 x 106, or from about 5 x 106to about 1 x 107) cells / cm3. In some cases, the enriched product can be enriched by at least 5-fold (e.g., at least 10-fold, at least 15-fold, at least 20-fold, at least 50-fold, or at least 100-fold) for a particular population of cells per mg of biological material as compared to the biological material prior to enrichment.

[0039] Any appropriate type of cells and other material can be included in an enriched end product generated using the devices and methods provided herein. For example, an enriched end product can contain ADSC, adipose-derived stromal cells (also known as adipose-derived adult stromal cells or adipose stromal cells), lipoblasts, pericytes, preadipocytes, processed lipoaspirate cells, endothelial cells, muscle cells (e.g., smooth muscle cells), fibroblasts, pericytes, perivascular cells, mural cells, immune cells (e.g., macrophages, monocytes, lymphocytes, T cells, and / or B cells), hormones, exosomes, purified adipose tissue, erythrocytes, and / or other stem cells (e.g., mesenchymal stem cells). In some cases, an enriched end product generated using the devices and methods provided herein can be a stromal vascular fraction. In general, a device provided herein can have a conduit (e.g., a tube) that extends from an inlet port to an outlet port. The inlet port can be designed to allow for the input of any appropriate biological material (e.g., fat tissue). An inlet port can be designed to have any appropriate configuration. For example, an inlet port of a device provided herein can be designed to connect to an outlet of a liposuction device so as to collect fat tissue being extracted from a mammal (e.g., a human). In some cases, an inlet port of a device provided herein can be designed to connect to a luer of a syringe containing biological material (e.g., tissue such as fat tissue) obtained from a mammal. The connection between an inlet port of a device provided herein and another device (e.g., a liposuction device or a syringe) can be any appropriate type of connection (e.g., a friction fit, force- fit, form-fit, and / or a material-fit connection such as a clamp, screw, snap, adhesive, or welded connection). The connection can be a detachable connection, a non-detachable connection, and / or a conditionally detachable connection. In some cases, a connection between an inlet port of a device provided herein and another device (e.g., a liposuction device or a syringe) capable of providing biological material obtained from a mammal can be designed to facilitate sterile transfer of the biological material into the conduit, such that the biological material is not exposed to contaminants in the environment outside the device. In some cases, a device provided herein can have an adapter that can be within the inlet port or within the conduit adjacent to the inlet port, or that can be positioned at the connection between the inlet port of the device and another device (e.g., a liposuction device or a syringe) from which biological material is to be transferred into the inlet port. In some cases, the adapter can be configured to, for example, facilitate flow of biological material into the inlet port. For example, an adapter can be configured to have a shape (e.g., a large, funnel-shaped geometry), one or more surface properties (e.g., microstructured surface, wettability, and / or applied coatings), an ability to adjust viscosity (e.g., lowering viscosity by having the ability to add water), and / or an ability to introduce internal and / or external forces or pressure differences to improve the input of a biological material into the device, if desired.

[0040] Any appropriate biological material can be inserted into a device (e.g., into a conduit of a device) provided herein. For example, biological material containing any appropriate type of tissue (e.g., adipose tissue, endothelial tissue, muscle tissue, bone marrow, blood, or umbilical cord tissue) or cells (e.g., adipocytes, endothelial cells, muscle cells, fibroblasts, pericytes, perivascular cells, mural cells, macrophages, monocytes, lymphocytes, T cells, B cells, adipose-derived stem cells, mesenchymal stem cells, adipose-derived regenerative cells, progenitor cells, preadipocytes, or erythrocytes) can be inserted into a device provided herein. In some cases, fat tissue can be inserted into a device provided herein. In some cases, bone marrow can be inserted into a device provided herein. In some cases, blood can be inserted into a device provided herein.

[0041] In some cases, the biological material can be combined with a fluid (e.g., sterile water or saline) before or during insertion into the inlet port of a device provided herein. Without being limited to any particular mechanism of action, the added fluid can modify (e.g., reduce) the viscosity of the biological material, which can facilitate passage of the biological material through the conduit of a device provided herein.

[0042] The biological material can be from any appropriate mammal. Examples of mammals from which biological material can be obtained for use in a device provided herein include, without limitation, humans, non-human primates (e.g., monkeys), dogs, cats, equines (e.g., horses, donkeys, camels, and zebras), bovines (e.g., cows, bison, and buffalo), pigs, sheep, mice, and rats. In some cases, the biological material can be from a human (e.g., a human in need of receiving stem cells to treat a medical condition such as osteoarthritis, bone / chondral disrepair, rheumatic diseases, muscle degeneration, facet joint syndrome, (rotator cuff) tendinopathy, chronic wounds, diabetic foot, hair loss, or multiple sclerosis). In some cases, the biological material can be from a non-human mammal (e.g., a horse having damage to its orthopedic musculoskeletal system, such as its articular cartilage and / or tendons, that can be improved by stem cell therapy). In general, treatments using the body’s own cells (e.g., stem cells) can alleviate a range of inflammatory and immune-mediated diseases in horses see, e.g., Pauwelyn and Glenn, Boehringer Ingelheim Pharma GmbH & Co. KG, www.boehringer- ingelheim.com / de / tiergesundheit / haustiere-und-pferde / stammzellen-therapien, 2024). When treating a horse, tendon and ligament injuries, osteoarthritis, lameness, degenerative joint changes, wounds, equine metabolic syndrome, diseases of the digestive tract, liver diseases, and / or neuromuscular diseases can be treated by removing a biological sample (e.g., adipose tissue) containing stem cells, processing the biological sample using a device provided herein, and returning the processed biological sample (e.g., a sample enriched for adipose-derived stem cells) to the horse to treat the disease(s).

[0043] In some cases, the biological material can be directly obtained from a mammal such that the biological material is collected directly from the mammal into an inlet port of a device provided herein. For example, when collecting fat tissue from a mammal (e.g., a human) during a liposuction procedure, an inlet port of a device provided herein can be connected to an outlet of the liposuction device. In some cases, when collecting a tissue sample, a bone marrow sample, or a blood sample from a mammal using a syringe connected to a needle (e.g., a biopsy needle or a needle typically used for blood collection), an inlet port of a device provided herein can be connected directly to a luer of the syringe.

[0044] In some cases, a biological material can enter a device provided herein in the same state it was in when it was extracted from the mammal. For example, a biological material can enter a device provided herein without being subjected to any purification. For example, a biological material can enter a device provided herein without the addition of any preservatives. For example, a biological material can enter a device provided herein without being diluted.

[0045] In some cases, a biological material can enter a device provided herein no more than about 5 minutes after being extracted from a mammal. For example, a biological material can enter a device provided herein within about 0 to about 5 minutes, about 0 to about 4 minutes, about 0 to about 3 minutes, about 0 to about 2 minutes, or about 0 to about 1 minute after being extracted from a mammal. For example, a biological material can enter a device provided herein instantaneously after being extracted from a mammal, such that the biological material enters the device directly and with no delay.

[0046] The outlet port can be designed to allow for the output of biological material after it has passed through the conduit and been processed as described herein. An outlet port can be designed to have any appropriate configuration. For example, an outlet port of a device provided herein can be designed to connect to a collection vessel. In some cases, an outlet port of a device provided herein can be designed to connect to an inlet of an administering device. For example, an outlet port of a device provided herein can be designed to connect to the barrel of a syringe so as to collect a processed biological material (e.g., an enriched population of cells) before transfer back into a mammal (e.g., the mammal from which the original biological material was obtained). In some cases, an outlet port of a device provided herein can be configured to deliver a processed biological material (e.g., an enriched population of cells) directly to a mammal (e.g., the mammal from which the original biological material was obtained). For example, an outlet port of a device provided herein can be configured so that processed biological material obtained can be applied directly by spatula and / or a spray nozzle to a surface to be treated. In some cases, an outlet port of a device provided herein can be configured so that processed biological material obtained can be fed directly back into the body via a needle-shaped end.

[0047] In some cases, an outlet port of a device provided herein can be designed to connect to a storage vessel (e.g., a tube or container) in which a processed biological material (e.g., an enriched population of cells) can be collected and stored. In some cases, the connection between an outlet port of a device provided herein and a collection vessel can be designed to facilitate sterile transfer of the biological material into the collection vessel, such that the biological material is not exposed to contaminants in the environment outside the device.

[0048] The conduit of a device provided herein can have any appropriate configuration. In some cases, the conduit of a device provided herein can be an elongate tube having an inlet port at one end and an outlet port at the other end. The conduit can have any appropriate length. For example, the length of a conduit of a device provided herein, from one end to the other, can be from about 2 cm to about 20 m (e.g., from about 2 cm to about 5 cm, from about 5 cm to about 10 cm, from about 10 cm to about 50 cm, from about 50 cm to about 100 cm, from about 100 cm to about 250 cm, from about 250 cm to about 500 cm, from about 500 cm to about 1 m, from about 1 m to about 2 m, from about 2 m to about 4 m, from about 4 m to about 6 m, from about 6 m to about 8 m, from about 8 m to about 10 m, or from about 10 m to about 20 m). The conduit of a device provided herein can have any appropriate cross-sectional shape and dimensions. In some cases, a conduit can have a circular cross-section or a substantially circular cross-section. In some cases, a conduit can have an oval crosssection, a substantially oval cross-section, or any cross-sectional area describable by a polygon (e.g., triangular, rectangular, etc.). In some cases, the cross-sectional shape of the conduit can vary along the length of the conduit. For example, a conduit can have one or more segments in which the cross-sectional shape is substantially circular, and one or more segments in which the cross-sectional shape is substantially oval. As described herein, for example, in some cases, a conduit can have a cross-sectional shape along its length that is substantially circular in the absence of any applied external force but is compressed to a more oval shape or elongated shape in locations or sections where outside force is applied (e.g., by a dispersing element).

[0049] The conduit of a device provided herein can have any appropriate inner diameter. For example, the inner diameter of a conduit can be from about 0.1 mm to about 20 mm (e.g., from about 0.1 mm to about 0.5 mm, from about 0.5 mm to about 1 mm, from about 1 to about 2 mm, from about 2 to about 3 mm, from about 3 to about 4 mm, from about 4 to about 5 mm, from about 5 to about 7.5 mm, from about 7.5 to about 10 mm, from about 10 to about 15 mm, or from about 15 mm to about 20 mm). In some cases, the conduit of a device provided herein can have a maximum inner diameter at any point along its length that is no greater than about 3 mm, no greater than about 5 mm, or no greater than about 10 mm. In some cases, the conduit of a device provided herein can have minimum inner diameter along the entirety of its length that is no less than about 2 mm, no less than about 1.5 mm, no less than about 1 mm, or no less than about 0.5 mm, except for one or more segments that can be designed to be used as dispersing elements to disperse the biological material into a more dispersed material.

[0050] In some cases, a conduit of a device provided herein can have an inner diameter that is constant along its entire length. In some cases, a conduit of a device provided herein can include two or more (e.g., two, three, four, five, or more than five) segments, where adjacent segments have different inner diameters. For example, a conduit of a device provided herein can have a first segment starting at the inlet port and a second segment ending at the outlet port, where the second segment has an inner diameter that is less than the inner diameter of the first segment. In some cases, a conduit of a device provided herein can have a first segment downstream of the inlet port having a first inner diameter and one or more internal segments along the length of the conduit having an inner diameter that is smaller than the first diameter. In some cases, such smaller inner diameter(s) can be designed to be used as dispersing elements to disperse the biological material into a more dispersed material. In some cases, a conduit of a device provided herein can have a first segment starting at the inlet port, one or more internal segments, and a last segment ending at the outlet port, where each segment from the inlet port to the outlet port has a progressively smaller inner diameter.

[0051] The conduit can be made from any appropriate material. In some cases, the conduit can be made from flexible tubing. In some cases, a conduit can include stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane (e.g., thermoplastic polyurethane), polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber (e.g., natural or synthetic rubber), or any combination thereof.

[0052] A device provided herein also can include one or more pumping elements configured to move biological material through the conduit. For example, a device provided herein can include one, two, three, four, five, or more than five pumping elements. The pumping element(s) can be engaged with the conduit to promote movement of biological material through the conduit such that the biological material moves away from an inlet port and toward an outlet port. Any appropriate type of pumping element can be included. For example, a pumping element can be a displacement pump (e.g., a reciprocating pump such as a diaphragm pump, or a rotary / continuous pump such as a peristaltic pump). In another example, a pumping element can be a direct lift device (e.g., a reciprocating direct lift device, or a rotary / continuous direct lift device). In another example, a pumping element can be a velocity pump (e.g., a reciprocating velocity pump or a rotary / continuous velocity pump), a buoyancy pump, an impulse pump, or a gravity device. In some cases, a plurality of different types of pumping elements (e.g., two or three different types of pumping elements) can be included.

[0053] The one or more pumping elements can be located at any appropriate position with respect to the conduit of a device provided herein. For example, a pumping element can be positioned to contact only an exterior portion of the conduit, such that the pumping element is not positioned within the conduit. In some cases, the entirety of the one or more pumping elements can be located outside the conduit, such that no part of any pumping element is located within the conduit. In some cases, a device provided herein can be designed so that no component of any pumping element is in contact with biological material within the conduit, whether the biological material is in its original form, its dispersed form, or its processed (e.g., enriched) form.

[0054] In some cases, a device provided herein can include a single pumping element, where the pumping element is located near the inlet port of the conduit. The action of the pumping element can move biological material within the conduit in a direction away from the inlet port and toward one or more dispersing elements, one or more separating elements, and an outlet port. In some cases, a device provided herein can include more than one pumping element (e.g., two, three, four, five, or more than five pumping elements). In some cases, all of the pumping elements can be located toward the inlet port end of the conduit, upstream of one or more dispersing elements and one or more separating elements. In some cases, the two or more pumping elements can be located along the length of the conduit (e.g., interspersed with one or more dispersing elements and one or more separating elements). In some cases, a device can be configured such that the conduit forms two or more loops (e.g., two, three, four, five, or more than five loops), where each loop passes through or is otherwise acted on by a pumping element. For example, each loop can be acted on by a separate pumping element, or two or more (e.g., all) loops can be acted on by the same pumping element.

[0055] A device provided herein also can include one or more dispersing elements. For example, a device provided herein can include one, two, three, four, five, six, seven, eight, nine, ten, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or more than 35 dispersing elements. The dispersing element(s) can be configured to break up biological material within the conduit, forming a more dispersed form of the biological material. Any appropriate type of dispersing element can be included. In some cases, a dispersing element can be an element that utilizes fluid mechanics to disperse biological material. For example, the conduit of a device provided herein can include one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or more than 35) bottlenecks or constrictions along its length that can reduce the open area of flow for the biological material in the conduit. As the biological material passes through the bottleneck(s) or constriction(s), the biological material can be broken up such that cells within the biological material are dissociated from other material and / or other cells. In some cases, a bottleneck or constriction can be a compressed portion of the conduit, where the portion is (or has been) compressed by an externally applied force. When a conduit includes two or more compressed portions, the compressed portions can be compressed in the same direction relative to one another, or in different directions relative to one another. For example, the compressed portions can be compressed in the same direction relative to one another such that when the conduit is viewed from the inlet port toward the outlet port (or vice versa), the compressed portions appear as flattened areas that line up with one another. Alternatively, the compressed portions can be compressed in different directions relative to one another, such that when the conduit is viewed from the inlet port toward the outlet port (or vice versa), the compressed portions appear as flattened areas that are at different angles relative to one another. In some cases, a bottleneck or a constriction can be a narrowed portion of the conduit material (e.g., a segment of tubing having a maximum internal diameter that is smaller than the maximum internal diameter of the adjacent upstream sections of tubing).

[0056] In some cases, a conduit can include two or more bottlenecks or constrictions, where each adjacent pair of bottlenecks or constrictions are separated by an intervening segment of the conduit. For example, each bottleneck or constriction can be from about 0.05 cm to about 3 cm in length (e.g., from about 0.05 to about 0.1 cm, from about 0.1 to about 0.5 cm, from about 0.5 to about 1 cm, from about 1 to about 2 cm, or from about 2 to about 3 cm in length). The intervening segments also can have any appropriate length. For example, each intervening segment can be from about 0.1 cm to about 20 cm in length (e.g., from about 0.1 to about 0.5 cm, from about 0.5 to about 1 cm, from about 1 to about 2 cm, from about 2 to about 4, from about 4 to about 6, from about 6 to about 8, from about 8 to about 10, from about 10 to about 15, or from about 15 to about 20 cm in length). In some cases, a device provided herein can include a conduit configured to form two or more loops (e.g., two, three, four, five, or more than five loops), where each loop includes three to ten (e.g., three to five, four to six, five to seven, six to eight, seven to nine, or eight to ten) bottlenecks or constrictions. In some cases, when a conduit forms two or more loops, each loop can pass through or otherwise interact with a pumping element, which can facilitate passage of biological material through the conduit and its bottlenecks or constrictions.

[0057] In some cases, one or more dispersing elements of a device provided herein can be a microstructure formed on an inner surface of the conduit. Examples of microstructures include, without limitation, microchannels, plate type microstructures, and housed-elements including of series of baffles made of, without limitation, metal, ceramics, glass, or plastic. The microstructures typically are less than 1 mm in size.

[0058] In some cases, a dispersing element can be non-integral to the conduit. For example, a dispersing element can include a rotor-stator system, an ultrasonic generator, a biochemical solution, or a chemical system. In some cases, a dispersing element can be an active mixer / stirrer (e.g., a stirrer, a mixing pump, and / or an ultrasonic generator), a passive mixer / stirrer (e.g., a flow mixer), a biochemical solution (e.g., one or more enzymes), or a chemical system (e.g., a solvent or surfactant). In some cases, such as when a dispersing element is an element such as a rotor-stator system or an ultrasonic generator, the dispersing element can be positioned to contact only an exterior surface of the conduit, such that it is not positioned within the conduit. In some cases, the entirety of the one or more dispersing elements can be located outside the conduit, such that no part of any dispersing element is located within the conduit. In some cases, a device provided herein can be designed so that no component of any dispersing element is in contact with biological material within the conduit, whether the biological material is in its original form, its dispersed form, or its processed (e.g., enriched) form. In some cases, a device provided herein can include a single dispersing element, where the dispersing element is located between at least one pumping element and an outlet port of the conduit. The action of the dispersing element can form a more dispersed form of the biological material within the conduit. In some cases, a device provided herein can include more than one dispersing element (e.g., two, three, four, five, or more than five dispersing elements). In some cases, all of the dispersing elements can be located at least somewhat centrally along the conduit, such that they are downstream of all of the one or more pumping elements and upstream of all of the one or more separating elements. In some cases, the two or more dispersing elements can be interspersed with two or more pumping elements and one or more separating elements.

[0059] In some cases, a device provided herein can have dispersing elements (e.g., constriction points) integrated directly into a pumping element (e.g., a peristaltic pump) configured to move cellular material through a conduit. A standard peristaltic pump typically has one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than then) fixed sliding shoes or rotatable rollers mounted on a pump impeller. The one or more sliding shoes or rollers can squeeze a conduit (e.g., tubing) associated with the pump at pinch points adjacent to the shoes or rollers, effectively sealing the conduit. Moreover, rotation of the pump impeller can cause the shoes or rollers to move material within the conduit forward through the device.

[0060] In some cases, a device provided herein can be configured to combine the conveying function of a pumping element (e.g., a peristaltic pump) with the dispersing elements (e.g., pins or other elements that can compress the conduit of a peristaltic pump to result in shearing at pinch points) in a single assembly. Such an arrangement can have advantages such as shortening the length of the conduit in the pumping element, which can reduce the volume of biological material required, and simplifying the machine by integrating the pumping and dispersing functions. It is to be noted that such a device might have a collision problem between moving rollers / sliding shoes and pinching pins, but such a collision problem can be avoided by, for example, using pins that are springmounted, using pins that are designed to be elastic (e.g., that are made, at least in part, of an elastic material such as rubber, silicone, or a thermoplastic elastomer such as a thermoplastic polyurethane or a thermoplastic polyolefin), using rollers or sliding shoes that are radially movable or elastic (e.g., that are made, at least in part, from an elastic material such as rubber or silicon), or any combination of these approaches to exert force against the tubing. For example, the action of one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than ten) spring-mounted elements to compress the conduit of a device provided herein (e.g., by pressing against an external surface of the conduit that is distal to the pump impeller) can be periodic and / or reversible, such that each element can press against the conduit for an appropriate period of time (e.g., about 0.5 second, about one second, about two to five seconds, or more than five seconds), and then release to allow the portion of the conduit previously compressed by the action of the element to return to a non-compressed configuration. When a device includes one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than ten) radially movable elements, the radially movable elements can reversibly exert force against an external surface of the conduit that is proximal to the pump impeller. The action of such radially movable elements to compress the conduit can be periodic and / or reversible, such that each element can press against the conduit for an appropriate period of time (e.g., about 0.5 second, about one second, about two to five seconds, or more than five seconds) to compress the conduit and aid in dispersal of a material contained therein, and also to convey the material through the conduit, and can then release to allow the portion of the conduit previously compressed by the action of the element to return to a non-compressed configuration. In some cases, a device provided herein that has radially movable also can include one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than ten) stationary elements that can press against an external surface of the conduit that is distal to the pump impeller, resulting in compressed portions (e.g., bottlenecks) in the conduit that aid in dispersing material contained therein. In some cases, a device provided herein can have a combination of radially movable elements and spring-mounted or elastic elements.

[0061] In some cases, the components (e.g., pins and / or other elements) that exert force against and compress the conduit can be constantly pressed against the conduit by a force that is, for example, generated mechanically (e.g., using pre-tensioned springs and / or using a belt or a thin, flexible metal sheet with protruding nubs), or generated using a fluid (e.g., a pneumatic or hydraulic fluid) and small pneumatic or hydraulic pistons. In some cases (e.g., depending on the material from which the conduit is made, the geometry of the conduit, and the desired level of compression), the distance between the pump rollers, sliding shoes, and / or pins at the moment when material within the conduit passes can be smaller than the minimum residual height of the squeezed conduit. In this case, collision can be passively avoided by elastic deformation or radial displacement of the pins and / or the rollers and / or sliding shoes. The required deformation energy can be provided by the pump motor, for example. Actively controlled pins and / or rollers and / or sliding shoes also can be used with an appropriate mechatronic system. The actuators in such a system can be mechanical, electrical, hydraulic, or pneumatic.

[0062] A device provided herein can include one or more separating elements. For example, a device provided herein can include one, two, three, four, five, or more than five separating elements. The one or more separating elements can be configured to separate cells within the dispersed biological material from other components of the biological material, forming an enriched population of cells. Any appropriate type of separating element can be included. In some cases, a separating element can form an enriched population of cells based on a particular characteristic of the enriched cells as compared to other cells or components within the biological material. For example, a separating element can separate a selected population of cells from other cells or components of a biological material based on the size, surface wettability, visual or optical reflective characteristics, density, inertia, magnetizability, or electrical mobility of the selected population.

[0063] In some cases, a separating element can include a filter. Any appropriate type of filter can be used. For example, a separating element can include a tangential flow filter (e.g., a hollow fiber filter, a diaphragm filter, or a filter having additively manufactured, integrated filter elements). In some cases, a filter can include filtration elements that are 3D printed structures (e.g., bars) integrated directly into the filter housing, where the structures define a corresponding pore size as a result of the distance between adjacent structures and the height of the channel. When a separating element includes a filter, the filter can have any appropriate pore size. For example, a separating element can include a filter having a pore size from about 1 pm to about 120 pm (e.g., from about 1 to about 10 pm, from about 10 to about 20 pm, from about 20 to about 30 pm, from about 30 to about 40 pm, from about 40 to about 60 pm, from about 60 to about 80 pm, from about 80 to about 100 pm, or from about 100 to about 120 pm). The pore size can be selected to remove unwanted cells and / or other components of a biological material from the cells and, if any, other material to be enriched. For example, in some cases, a filter having a pore size from about 50 to about 100 pm (e.g., about 80 pm) can be used to remove cells and other items larger than about 80 pm (including, e.g., adipose cells) from a dispersed form of a biological material. In some cases, a filter having a pore size from about 5 to about 15 pm (e.g., about 10 pm) can be used to remove cells and other items smaller than about 10 pm (e.g., fat globules and erythrocytes) from a dispersed form of a biological material. In some cases, a device provided herein can include two or more separating elements, where each separating element includes a filter, and where the filters have different pore sizes. For example, a device provided herein can include one or more filters as separating elements having a pore size from about 50 to about 100 pm (e.g., about 80 pm) to remove items larger than about 50 to about 100 pm (e.g., about 80 pm) such as adipose cells from a dispersed form of a biological material with the resulting items having a size smaller than the pore size advancing to one or more additional filters as separating elements having a pore size from about 5 to about 15 pm (e.g., about 10 pm) to remove items smaller than about 5 to about 15 pm (e.g., about 10 pm) such as fat globules and erythrocytes from a dispersed form of a biological material with the resulting items having a size larger than the pore size of those additional filters advancing toward that outlet port. In some cases, a device provided herein can include one or more filters as separating elements having a pore size from about 5 to about 15 pm (e.g., about 10 pm) to remove items smaller than about 5 to about 15 pm (e.g., about 10 pm) such as fat globules and erythrocytes from a dispersed form of a biological material with the resulting items having a size larger than the pore size advancing to one or more additional filters as separating elements having a pore size from about 50 to about 100 pm (e.g., about 80 pm) to remove items larger than about 50 to about 100 pm (e.g., about 80 pm) such as adipose cells from a dispersed form of a biological material with the resulting items having a size smaller than the pore size of those additional filters advancing toward that outlet port. The one or more separating elements can be located at any appropriate position with respect to the conduit of a device provided herein. For example, since the one or more separating elements typically contact the biological material from which an enriched cell population is to be separated, the one or more separating elements can be positioned within the conduit of a device provided herein. When two or more separating elements are included in a device provided herein, the separating elements can be positioned in any appropriate configuration relative to one another. For example, two or more separating elements (e.g., two or more filters) can be present in a device provided herein in an axial arrangement, in a radial arrangement, in a parallel arrangement, in a serial arrangement, or in a combination of such arrangements (e.g., in a serial and parallel arrangement).

[0064] In some cases, a device provided herein can include a single separating element, where the separating element is located downstream of the at least one pumping element and the at least one dispersing element. In some cases, a device provided herein can include more than one separating element (e.g., two, three, four, five, or more than five separating elements). In some cases, all of the separating elements can be located downstream of all of the one or more pumping elements and all of the one or more separating elements. In some cases, the two or more separating elements can be interspersed with two or more dispersing elements.

[0065] In some cases (e.g., when a device is configured to separate stem cells), a device provided herein can include at least a two-stage filter. Filters having more than one stage (e.g., two, three, four, or more than four stages) can be generated using any appropriate method, including additive manufacturing (also referred to as 3D printing). Devices that include two or more filtration stages can be built into one or more units. For example, a device can include a single separating element (i.e., a single unit) that includes two or more filter stages. Each of the one or more units can be connected to the conduit, such that the flow of material from the conduit can be supplied to and subsequently discharged from the filter stages. Each filter stage can include, for example, one or more membranes, a multi-part housing, one or more flow channels, and / or one or more vent valves. In some cases, a device provided herein can include a first filter stage having a flow-through filter, and a second, fine filter stage having a hollow fiber filter or a tangential flow filter (e.g., with a membrane surface and / or 3D-printed bars) to filter out most of the particles that are too fine as permeate. Using additive manufacturing, such filter stages can be produced in a single manufacturing process. Moreover, additional functions (e.g., one or more fastening elements, collecting vessels, pressure control elements, vents, and / or elements for controlling the flow of material) can be integrated at the same time.

[0066] In some cases, a device provided herein can include one or more (e.g., one, two, three, four, five, or more than five) holding chambers. The one or more holding chambers can be in fluid communication with the conduit, and can be configured to accumulate and contain biological material (e.g., biological material in its original form, biological material in a more dispersed form, and / or biological material in an enriched form). For example, a device provided herein can include one or more holding chambers located along a conduit between an inlet port and a first dispersing element, where the one or more holding chambers are configured to accumulate and contain biological material before it is dispersed. In some cases, a device provided herein can include one or more holding chambers located along a conduit between a first dispersing element and a first separating element, where the one or more holding chambers are configured to accumulate and contain a dispersed form of biological material. In some cases, a device provided herein can include one or more holding chambers located along a conduit between a first separating element and an outlet port, where the one or more holding chambers are configured to accumulate and contain an enriched population of cells.

[0067] In some cases, the one or more holding chambers can be connected to the conduit of a device provided herein by a force-fit, a form-fit, and / or a material-fit connection such as a clamp, friction, screw, snap, adhesive, and / or welded connection. In some cases, the connection can be a detachable connection, a non-detachable connection, or a conditionally detachable connection. In some cases, the one or more holding chambers can be used to smooth the flow of material and / or to store (e.g., store temporarily) the material for, during, or after processing. The movement of the material can be caused by internal and / or external forces and / or pressure differences.

[0068] When one or more holding chambers are included in a device provided herein, the holding chambers can have any appropriate capacity. For example, a holding chamber can be configured to contain from about 1 mL to about 3000 mL of biological material (e.g., from about 1 mL to about 10 mL, from about 10 mL to about 50 mL, from about 50 mL to about 100 mL, from about 100 mL to about 300 mL, from about 300 mL to about 500 mL, from about 500 mL to about 1000 mL, from about 1000 mL to about 2000 mL< or from about 2000 mL to about 3000 mL). In some cases, when two or more holding chambers are included, the holding chambers can have the same capacity. In some cases, when two or more holding chambers are included, the holding chambers can have different capacities.

[0069] In some cases, a device provided herein can include one or more (e.g., one, two, three, four, five, or more than five) inflow ports in fluid communication with the conduit. The one or more inflow ports can be configured to add a fluid (e.g., water, saline, or a solution containing one or more cytokines, growth factors, vitamins, hormones, phosphate-buffered saline, culture fluid, blood, a solution containing one or more components of fat tissue or cells, and / or drug active ingredients such as acetylsalicylic acid) to biological material contained within the conduit. When one or more inflow ports are included, they can be located at any appropriate position along the conduit between the inlet port and the outlet port. For example, a device provided herein can include one or more inflow ports fluidly connected to the conduit between the inlet port and a first dispersing element, and / or one or more inflow ports fluidly connected to the conduit between a first dispersing element and a first separating element, and / or one or more inflow ports fluidly connected to the conduit between a first separating element and an outlet port. In some cases, a device provided herein can lack inflow ports along the conduit between the inlet port and the outlet port.

[0070] Representative embodiments of devices provided herein are illustrated in Figures 1-6 and Figures 10-14E. With reference to Figure 1, device 100 can be designed to include a conduit 120 having an inlet port 110 and an outlet port 160. Device 100 also can include a pumping element 130, a dispersing element 140, and a separating element 150. The pumping, dispersing, and separating elements of device 100 can be located at any appropriate position relative to one another. As illustrated in the representative example of Figure 1, the pumping element can be proximal to the inlet port, the dispersing element can be downstream from the pumping element, and the separating element can be downstream of the dispersing element.

[0071] With reference to Figure 2, device 200 can be designed to include more than one pumping element, more than one dispersing element, and / or more than one separating element. A representative example of such an embodiment is illustrated in Figure 2. As shown in Figure 2, device 200 can have three pumping elements (130, 132, and 134), two dispersing elements (140 and 142), and four separating elements (150, 152, 154, and 156). The pumping, dispersing, and separating elements of device 200 can be located at any appropriate position relative to one another. As illustrated in the representative example of Figure 2, the pumping elements can be proximal to the inlet port, the dispersing elements can be downstream from the pumping elements, and the separating elements can be downstream of the dispersing elements.

[0072] With reference to Figure 3, device 300 can be designed to include one or more holding chambers in fluid communication with conduit 120. A representative example of such an embodiment is illustrated in Figure 3. As shown in Figure 3, device 300 can have first holding chamber 170, second holding chamber 172, and third holding chamber 174. The one or more holding chambers can be located at any appropriate position along conduit 120. As illustrated in the representative example of Figure 3, first holding chamber 170 can be located between inlet port 110 and pumping element 130, second holding chamber 172 can be located between dispersing element 140 and separating element 150, and third holding chamber 174 can be located between separating element 150 and outlet port 160.

[0073] With reference to Figure 4, device 400 can be designed to include one or more inflow ports in fluid communication with conduit 120. A representative example of such an embodiment is illustrated in Figure 4. As shown in Figure 4, device 400 can have first inflow port 180 and second inflow port 185. The one or more inflow ports can be located at any appropriate position along conduit 120. As illustrated in the representative example of Figure 4, first inflow port 180 can be located between pumping element 130 and dispersing element 140, and second inflow port 185 can be located between dispersing element 140 and separating element 150.

[0074] With reference to Figure 5, device 500 can be designed to include a conduit that varies in diameter along its length. A representative example of such an embodiment is illustrated in Figure 5. As shown in Figure 5, conduit 120 of device 500 can include first section 190 and second section 195, where first section 190 is upstream of second section 195, and where the diameter of second section 195 is smaller than the diameter of first section 190.

[0075] With reference to Figures 6A and 6B, device 600 can include conduit 610 having inlet port 615 and outlet port 690, pumping element 660, and separating elements 680 and 685. Conduit 610 can be flexible and can be configured to form a series of loops 620, 630, 640, and 650. Each loop of conduit 610 can include several compressed regions (e.g., compressed regions 621, 622, 623, 624, and 625 (with 622-624 shown as 62x) of loop 620, compressed regions 631, 632, 633, 634, and 635 of loop 630, compressed regions 641, 642, 643, 644, and 645 of loop 640, and compressed regions 651, 652, 653, 654, and 655 (with 652-654 shown as 65x)of loop 650). Pumping element 660 can be a peristaltic pump with a series of pump wheels 662, 664, 666, and 668 fixed on axel 670. Because pump wheels 662, 664, 666, and 668 are fixed on axel 670, the pump wheels can rotate about axel 670 at the same speed relative to one another. Loop 620 can be positioned on pump wheel 662, loop 630 can be positioned on pump wheel 664, loop 640 can be positioned on pump wheel 666, and loop 650 can be positioned on pump wheel 668, such that the rotating action of the pump wheels when axel 670 rotates can convey biological material through conduit 610. System 600 can further include separating elements 680 and 685, which can be filters with differing pore sizes. For example, separating element 680 can be a filter having a pore size effective to remove particles (e.g., fat particles) that are larger than the desired cells, and separating element 685 can be a filter having a pore size effective to remove particles (e.g., blood particles) that are smaller than the desired cells.

[0076] With regard to Figure 10, device 1000 can include peristaltic pump housing 1040, which can be configured to contain pump impeller 1030, a flexible hollow conduit (e.g., tubing 1010) that can contain a material to be conveyed, and one or more fixed sliding shoes or rotatable rollers (e.g., rollers 1020, 1022, 1024, and 1026) mounted on pump impeller 1030. Rollers 1020, 1022, 1024, and 1026 can effectively squeeze tubing 1010 at pinch points adjacent to rollers 1020, 1022, 1024, and 1026 (e.g., pinch points 1050, 1052, and 1054), effectively sealing the tubing. Moreover, the rotation of pump impeller 1030 can cause rollers 1020, 1022, 1024, and 1026 to rotate at the pinch points, which can move material within tubing 1010 forward through the tubing.

[0077] With regard to Figure 11, device 1060 can combine the conveying function of a peristaltic pump with dispersing elements that include elastic and / or spring-mounted pins (e.g., spring-mounted pins 1061, 1062, 1063, 1064, 1065, 1066, and 1067). With regard to Figure 12, device 1070 can combine the conveying function of a peristaltic pump with dispersing elements that are radially movable (e.g., radially movable and / or elastic rollers

[0078] 1071, 1072, 1073, and 1074). Device 1070 also can include stationary dispersing elements (e.g., pins 1081, 1082, 1083, 1084, 1085, 1086, and 1087) that can press against an external surface of tubing 1010 that is distal to the pump impeller, resulting in compressed portions (e.g., bottlenecks) in tubing 1010 that aid in dispersing material contained within the tubing. With regard to Figure 13, device 1090 can combine the conveying function of a peristaltic pump with dispersing elements that include a combination of elastic and / or spring-mounted pins (e.g., pins 1061, 1062, 1063, 1064, 1065, 1066, and 1067) and elastic and / or radially movable elements (e.g., rollers 1071,

[0079] 1072, 1073, and 1074). In some cases, as depicted in Figure 13, a device provided herein can have a combination of radially movable elements (e.g., radially movable rollers 1071, 1072, 1073, and 1074) and spring-mounted elements (e.g., spring-mounted pins 1061, 1062, 1063, 1064, 1065, 1066, and 1067).

[0080] Figure 14 shows an embodiment of an entire separation element (a filter unit), while Figures 15 to 18 show details of several embodiments of separation elements. With regard to Figure 14, the dotted arrows depict the flow of biological material through separation element 1100 having first filter stage 1130 and second filter stage 1160. Separation element 1100 can be connected to a conduit via inlet connection 1110. Biological material flowing into first filter stage 1130 via inlet connection 1110 can pass through a channel in base plate 1115 and then into first filter stage 1130. As depicted in Figure 14, the direction of flow through the elements of separation unit 1100 is upwards or sideways. Without being bound by a particular mechanism of action, movement of biological material in an upward or sideways direction can facilitate the removal of air bubbles from the material.

[0081] In some cases, first filter stage 1130 can be a flow-through filtration unit having two or more (e.g., two, three, four, five, or more than five) stacked filters (e.g., filter disks 1131, 1132, 1133, and 1134). In first filter stage 1130, the flow of material can be deflected radially outward from a point of entry from base plate 1115. The flow can then progress through stacked filter disks 1131, 1132, 1133, and 1134 from the outside inward, into collection channel 1136 and then upward into second filter stage 1160.

[0082] As depicted in Figure 14, biological material can flow upward from first filter stage 1130 to enter second filter stage 1160 through a bottom opening. The material can then flow tangentially past the filter surfaces (e.g., filter membranes 1162 and 1164). The permeate can then be collected externally in a ring channel and discharged from second filter stage 1160 through outlet 1168.

[0083] Device 1100 also can have vertical column 1170 in fluid connection with horizontal conduit 1175, rinse collecting receptacle 1180, and retentate collecting receptacle 1190. The concentrated retentate from second filter stage 1160 can be fed upward through vertical column 1170, which can equalize the surface tension at small (e.g., about 10 pm in size) pores in the filter stage so that the liquid can pass through the pores, and the retentate can then move into horizontal conduit 1175 toward rinse collecting receptacle 1180 and retentate collecting receptacle 1190. When rinse collecting receptacle 1180 is full, the material flow of the retentate can overcome a height offset in horizontal conduit 1175 and reach retentate collecting receptacle 1190. See, the blown up view in Figure 18 for additional details. The prepared biological material (the retentate) can then be removed from retentate collecting receptacle 1190 through opening 1195. For example, the retentate within retentate collecting receptacle 1190 can be drawn up through opening 1195 with a syringe and then delivered (either directly or at a later time) to the mammal from which the biological sample was originally obtained.

[0084] Figures 15 and 16 depict two different design options for flow-through filters in first filter stage 1130. Again, the dotted arrows depict the flow of biological material through the filters. With regard to Figure 15, filter 1131 can have filter housing 1140 that contains bottom disk 1142, one or more connectors (e.g., connectors 1144) for connection to another filter, and a plurality of radial separators (e.g., radial separators 1144). A biological material can flow past radially arranged separators 1144. As depicted in Figure 15, radial separators can be positioned closer to one another toward the center of filter 1131, such that the distance between adjacent radial separators becomes smaller toward the center of filter 1131, and particles that are too large to pass through filter 1131 are gradually held back, while the smaller particles pass through first filter unit 1130.

[0085] With regard to Figure 16, filter 1132 can have filter housing 1140 that contains bottom disk 1142, one or more connectors (e.g., connectors 1144) for connection to another filter, and a plurality of concentric rings (e.g., rings 1148) having openings therethrough for the passage of material, such that the concentric rings function as an element for separation in the flow-through filtration. A biological material can flow through concentric rings 1148. As depicted in Figure 16, the openings though concentric rings for the passage of biological material can become progressively smaller from the outside of filter 1132 toward the center of filter 1132, such that particles that are too large to pass through the openings in concentric rings 1148 are gradually held back, while the smaller particles pass through first filter unit 1130.

[0086] Figure 17 illustrates a representative structure of second filter stage 1160. With respect to Figure 17, second filter stage 1160 can have central opening 1163, filter membranes 1162 and 1164, central channel 1165, outer ring channel 1167, outlet 1168, and opening 1169. Biological material can enter second filter stage 1160 through central opening 1163 and then flow tangentially past filter membranes 1162 and 1164. Due to hydrostatic pressure of the water column for the retentate (depicted in Figure 14), a large portion of the liquid material is pressed through small pores in filter membranes 1162 and 1164 as permeate, and guided toward outlet 1168 in outer ring channel 1167. The retentate is directed upward at an end of central channel 1165 through opening 1169, in the direction of vertical column 1170 (depicted in Figure 14).

[0087] Figure 18 depicts how a saline or water solution used for rinsing purposes can be separated from desired retentate material by vertical height offset 1178 in horizontal conduit 1175. Only when rinse collecting receptacle 1180 is filled can the processed biological material enter retentate collecting receptacle 1190. The retentate can be temporarily stored in receptacle 1190 and then removed through opening 1195 using, for example, a syringe.

[0088] In general, a device provided herein can initially be filled with a sterile fluid (e.g., saline or sterile water) via the inlet of the conduit, and can pass through the device, through inlet connection 1110, through separation element 1100, and up through vertical column 1170, in order to remove air from the device. The fluid can then be removed (e.g., pushed out of) the device by biological material that is placed in the device via the inlet of the conduit. Most of the sterile fluid (e.g., saline or water) can exit the filter housing via outlet 1168 (see, e.g., Figure 14). The remaining fluid can then be collected in container 1180. The size of the collecting container for the rinsing fluid and the height of vertical height offset 1178 can then ensure that only the desired concentrated material (e.g., the stem cells) reaches container 1190.

[0089] This document also provides methods for making a device provided herein. Any appropriate techniques can be used to make a device provided herein. In some cases, a device provide herein can be made by positioning one or more separating elements (e.g., filters) into a conduit (e.g., an appropriate length of flexible tubing). The one or more separating elements can be positioned into the conduit under sterile conditions, or the device can be sterilized after the separating element(s) have been positioned into the conduit. In some cases, a first tubing section containing the inlet port can be attached to the inflow end of a separating element (e.g., a filter element) and a second tubing section can be attached to the outflow end of the separating element (e.g., the filter element). In some cases, the filter element can be designed to allow biological material that is larger than a target size, at the target size (e.g., from about 8 to about 80 gm), and smaller than the target size to enter, while excluding (or filtering out) a majority (e.g., at least 60, 70, 80, 90, or 95 percent) of the larger and smaller items, thereby allowing material within the target size to exit and proceed toward the outlet port. In some cases, additive manufacturing technology can be used to generate the filter structures (e.g., membranes with pores) used in the devices provided herein. For example, a membrane can be 3D printed vertically, or thin filaments can be laid down horizontally at desired distance from one another to effectively yield pores having sizes that are defined by the distance between adjacent filaments. A method for making a device can further include positioning the conduit in contact with one or more pumping elements and one or more dispersing elements. In some cases, one or more inflow ports and / or one or more holding chambers can be coupled to the conduit such that they are in fluid communication with the conduit. The one or more inflow ports and / or holding chambers can be coupled to the conduit under sterile conditions, or the device can be sterilized after the one or more inflow ports and / or holding chambers have been coupled to the conduit.

[0090] A device provided herein can be made to have a first set of items such as a power supply, a control unit, material handling items, and fastening items and a second set of items that can be sterile and disposable. The device can be constructed such that biological material only comes into direct contact with the disposable set of items such that the feeding, conveying, processing (e.g., dispersing and separating), optional storage, and feeding out takes place within a sterile environment. In some cases, the device can ensure that biological material flows from an inlet port through the conduit and its components to an outlet port while the desired biological component is correspondingly concentrated.

[0091] The individual components of the disposable set of items (e.g., the ports, containers, conveying elements, dispersing elements, and separating elements) can be assembled manually at the point-of-care to a conduit and then fixed to a device or can be fixed to a device as a fully assembled, prefabricated set including conduit. Examples of appropriate types of connections for manual assembly include, without limitation, force- fit connections, form-fit connections, and / or material-fit connections such as clamp, friction, screw, snap, adhesive, and / or welded connection. Such connections can be detachable, non-detachable, or conditionally detachable connections.

[0092] In some cases, for reasons of hygiene, a completely assembled, prefabricated set of disposable items can be inserted into a machine containing the first set of items and can be connected sterilely to the same donor / recipient or to a donor and separate recipient of the biological material in, for example, a closed system.

[0093] This document also provides methods for using a device provided herein to generate an enriched population of cells from biological material obtained from a mammal (e.g., a human). In some cases, a method provided herein can include inserting biological material from a mammal (e.g., a human) into an inlet port of a device provided herein such that the biological material can access the conduit of the device, actuating a pumping element of the device to move the biological material within the conduit in a direction away from the inlet port and toward an outlet port of the conduit, actuating a dispersing element of the device to form a dispersed form of the biological material within the conduit, and allowing the dispersed form of the biological material to contact a separating element within the conduit, to form an enriched population of cells from the dispersed form of the biological material.

[0094] In addition to the steps listed above, in some cases, a method provided herein can include one or more optional steps. For example, a method can include obtaining the biological material from the mammal. It is to be noted that in some cases, obtaining biological material from a mammal and inserting the biological material into an inlet port of a device provided herein can be accomplished in a single step, such that the biological material is inserted directly from the mammal into the inlet port. For example, biological material (e.g., fat tissue) can be obtained using a liposuction device and passed directly from the liposuction device into the conduit of a device provided herein. In some cases, biological material from a mammal can be obtained in a receptacle (e.g., a syringe or a biopsy needle) and then passed from the receptacle into the inlet port and conduit of a device provided herein.

[0095] In some cases, a user can place the second set of sterile, disposable items into a device provided herein, and can fix the second set in the area of the filter housing. The user can then place the conduit in the pumping element (e.g., the peristaltic pump). The entire disposable set can then be rinsed (e.g., with saline) to replace air contained therein with saline. The device can then be connected to a source of biological material (e.g., via inlet 110 depicted in Figures 1-5) and the collection containers can be connected accordingly. In some cases, to ensure a high level of product safety in terms of batch tracking and preventing product piracy, a device provided herein can automatically recognize and record an inserted disposable set if the device is equipped with the appropriate equipment and has network access; such a device also can query the manufacturer’s approval (e.g., to match a barcode or RFID chip on one or more components of the disposable set and match the barcode or RFID chip with the manufacturer’s database) and then set the machine parameters accordingly. This also can ensure that each disposable set is only used once.

[0096] Any appropriate type of biological material can be obtained and placed into a device provided herein. Suitable types of biological material include, without limitation, fat tissue, blood, bone marrow, and muscle tissue. Further, any appropriate amount of biological material can be placed into the conduit of a device provided herein. For example, from about 0.01 L to about 5 L (e.g., from about 0.01 L to about 0.05 L, from about 0.05 L. to about 0.1 L, from about 0.1 L to about 0.5 L, from about 0.5 L to about 1 L, from about 1 L to about 2 L, from about 2 L to about 3 L, from about 3 L to about 4 L, or from about 4 L to about 5 L) of biological material can be inserted into the conduit of a device provided herein and processed to generate an enriched population of cells.

[0097] In some cases, a method provided herein can include administering an enriched product containing an enriched population of cells back to the mammal from which the biological material was obtained. Any appropriate amount of an enriched product can be administered. For example, a method provided herein can include administering from about 1 mL to about 500 mL (e.g., from about 1 mL to about 10 mL, from about 10 mL to about 25 mL, from about 25 mL to about 50 mL, from about 50 mL to about 100 mL, from about 100 mL to about 250 mL, or from about 250 mL to about 500 mL) of an enriched product to a mammal. The enriched product can be administered by any appropriate route (e.g., by direct injection into a particular organ or tissue, by intravenous injection, by open surgery, by minimally invasive surgery such as arthroscopy or endoscopy, by oral application, or by superficial (topical) application).

[0098] In some cases, a method can optionally include adding a fluid to biological material contained within the conduit. For example, a method can include introducing a fluid (e.g., water, saline, or a solution containing one or more cytokines, growth factors, or other additives) into an inflow port of a device provided herein. The fluid can be added at any appropriate step of a method provided herein. For example, a fluid can be combined with biological material before it is inserted into the inlet port, or a fluid can be introduced into the conduit of a device before or during dispersal of biological material contained therein, after dispersal but before separation of biological material contained therein, or during separation of the biological material to generate an enriched product. Any appropriate amount of a fluid can be introduced and combined with biological material. For example, from about 0.1 mL to about 2 mL (e.g., from about 0.1 mL to about 0.2 mL, from about 0.2 mL to about 0.3 mL, from about 0.3 mL to about 0.5 mL, from about 0.5 mL to about 0.75 mL, from about 0.75 mL to about 1 mL, from about 1 mL to about 1.5 mL, or from about 1.5 mL to about 2 mL) of fluid for every gram or mL of biological material can be introduced.

[0099] In some cases, a method provided herein can include collecting a biological material, a more dispersed form of the biological material, and / or an enriched population of cells in one or more holding chambers coupled to the conduit. The movement of biological material or processed biological material can be caused by internal and / or external forces and / or pressure differences.

[0100] In some cases, a method provided herein can include generating an enriched population of cells of from biological material obtained from a mammal and then returning the enriched population of cells to the mammal. For example, the biological material obtained from the mammal can be passed from the mammal, through a device provided herein, and - in its enriched form - back into the mammal. In some cases, the biological material obtained from the mammal can be passed from the mammal, through a device provided herein, and back into the mammal - in its enriched form - directly, such that the enriched population of cells is not exposed to open air and / or does not leave the surgical or clinical room where the mammal is located. In some cases, the biological material obtained from the mammal can be passed from the mammal, through a device provided herein, and back into the mammal - in its enriched form - directly, such that the enriched population of cells is not polluted.

[0101] In some cases, a method provided herein can include obtaining a biological material from a mammal, processing the biological material through a device provided herein to generate an enriched cell population from the biological material, and reintroducing the enriched cell population into the same mammal within about 5 to about 30 minutes. For example, biological material can be obtained from a mammal and processed through a device provided herein to generate an enriched cell population, and the enriched cell population can be re-introduced into the same mammal within about 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 15 minutes to about 20 minutes, about 20 minutes to about 25 minutes, or about 25 minutes to about 30 minutes.

[0102] In some cases, a method provided herein can include obtaining a biological material from a mammal, processing the biological material through a device provided herein to generate an enriched cell population from the biological material, and reintroducing the enriched cell population into the same mammal, where the method does not include a culturing step. For example, biological material can be obtained from a mammal and processed through a device provided herein to generate an enriched cell population, and the enriched cell population can be re-introduced into the same mammal without culturing the biological material or the enriched cell population.

[0103] With reference to Figure 7, for example, a method 700 provided herein can include placing biological material from a mammal into a conduit of a device provided herein (710), pumping the biological material through the conduit (720), dispersing the biological material within the conduit (730), separating the dispersed biological material within the conduit to generate an enriched population of cells (740), and collecting the enriched population of cells from the conduit (750). In some cases, the method optionally can include collecting biological material from a mammal (705) and / or administering the collected cells to the mammal (760). It is to be noted that steps within the method can be carried out concurrently. For example, pumping step 720 can be carried out concurrently with other steps of the method. For example, pumping step 720 can be continuously occurring, such that the biological material is being pumped through the conduit while dispersing step 730, separating step 740, and / or collecting step 750 are carried out. In addition, it is to be noted that dispersing step 730, separating step 740, and collecting step 750 can be carried out concurrently at different points along the conduit, such that different portions of the biological material are concurrently subjected to different steps of the method, depending upon their position within the conduit. Further, it is to be noted that dispersing step 730, separating step 740, and / or collecting step 750 can be repeated two or more times during execution of the method. For example, the enriched population of cells can be subjected to one or more further rounds of dispersing step 730 and / or one or more further rounds of separating step 740 before being collected.

[0104] In some cases, a method provided herein can include one or more steps in which the biological material is transferred to a holding chamber. With reference to Figure 8, for example, a method 800 provided herein can include placing biological material from a mammal into a conduit of a device provided herein (810), pumping the biological material through the conduit (820), transferring the biological material to a holding chamber (823), returning the biological material to the conduit (828), dispersing the biological material within the conduit (830), transferring the dispersed biological material to a holding chamber (833), returning the dispersed biological material to the conduit (838), separating the dispersed biological material within the conduit to generate an enriched population of cells (840), and collecting the enriched population of cells from the conduit (850). In some cases, the method optionally can include collecting biological material from a mammal (805) and / or administering the collected cells to the mammal (860). Again, it is to be noted that steps within the method can be carried out concurrently. For example, pumping step 820 can be carried out concurrently with other steps of the method. For example, pumping step 820 can be continuously occurring, such that the biological material is being pumped through the conduit while dispersing step 830, separating step 840, and / or collecting step 850 are carried out. In addition, it is to be noted that dispersing step 830, separating step 840, and collecting step 850 can be carried out concurrently at different points along the conduit, such that different portions of the biological material are concurrently subjected to different steps of the method, depending upon their position within the conduit. Further, it is to be noted that dispersing step 830, separating step 840, and / or collecting step 850 can be repeated two or more times during execution of the method. For example, the enriched population of cells can be subjected to one or more further rounds of dispersing step 830 and / or one or more further rounds of separating step 840 before being collected.

[0105] In some cases, a method provided herein can include one or more steps in which a fluid is added to the biological material. A fluid can be added at any appropriate step, including prior to or during pumping, prior to or during dispersing, prior to or during separation, prior to or during collection, or any combination thereof. With reference to Figure 9, for example, a method 900 provided herein can include placing biological material from a mammal into a conduit of a device provided herein (910), adding a fluid (e.g., water or saline) to the biological material (915), pumping the biological material through the conduit (920), adding a fluid (e.g., water or saline) to the pumped biological material (925), dispersing the biological material within the conduit (930), adding a fluid to the dispersed biological material (935), separating the dispersed biological material within the conduit to generate an enriched population of cells (940), and collecting the enriched population of cells from the conduit (950). In some cases, the method optionally can include collecting biological material from a mammal (905) and / or administering the collected cells to the mammal (960). In addition, it is to be noted that steps within the method can be carried out concurrently. For example, pumping step 920 can be carried out concurrently with other steps of the method. For example, pumping step 920 can be continuously occurring, such that the biological material is being pumped through the conduit while dispersing step 930, separating step 940, and / or collecting step 950 are carried out. In addition, it is to be noted that dispersing step 930, separating step 940, and collecting step 950 can be carried out concurrently at different points along the conduit, such that different portions of the biological material are concurrently subjected to different steps of the method, depending upon their position within the conduit. Further, it is to be noted that dispersing step 930, separating step 940, and / or collecting step 950 can be repeated two or more times during execution of the method. For example, the enriched population of cells can be subjected to one or more further rounds of dispersing step 930 and / or one or more further rounds of separating step 940 before being collected.

[0106] Exemplary Embodiments

[0107] Embodiment l is a device for obtaining an enriched population of cells from biological material obtained from a mammal, wherein the device comprises: (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the enriched population from the conduit, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of cells from the dispersed form of the biological material, wherein the enriched population of cells comprises a greater number of cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material.

[0108] Embodiment 2 is the device of embodiment 1, wherein the enriched population of cells is an enriched population of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells.

[0109] Embodiment 3 is the device of embodiment 1 or embodiment 2, wherein the cells are adipose-derived stem cells.

[0110] Embodiment 4 is the device of any one of embodiments 1-3, wherein the biological material comprises fat tissue.

[0111] Embodiment 5 is the device of any one of embodiments 1-4, wherein the mammal is a human.

[0112] Embodiment 6 is the device of any one of embodiments 1-4, wherein the mammal is a horse. Embodiment 7 is the device of any one of embodiments 1-6, wherein the conduit comprises flexible tubing.

[0113] Embodiment 8 is the device of any one of embodiments 1-7, wherein the conduit is from about 20 cm to about 10 m in length.

[0114] Embodiment 9 is the device of any one of embodiments 1-8, wherein the conduit comprises at least one section having an inner diameter of about 1 mm to 5 mm.

[0115] Embodiment 10 is the device of any one of embodiments 1-9, wherein a maximum inner diameter along any point of the conduit is no greater than 20 mm.

[0116] Embodiment 11 is the device of any one of embodiments 1-10, wherein a minimum inner diameter along any point of the conduit is no less than 0.1 mm.

[0117] Embodiment 12 is the device of any one of embodiments 1-11, wherein the conduit comprises stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof.

[0118] Embodiment 13 is the device of any one of embodiments 1-12, wherein the inlet port is configured to attach to an outlet of a liposuction device.

[0119] Embodiment 14 is the device of embodiment 13, further comprising an adaptor configured to attach the inlet port to the liposuction device.

[0120] Embodiment 15 is the device of any one of embodiments 1-12, wherein the inlet port is configured to collect the biological material directly from the mammal.

[0121] Embodiment 16 is the device of any one of embodiments 1-15, wherein the outlet port is configured to attach to an inlet of an administering device.

[0122] Embodiment 17 is the device of any one of embodiments 1-15, wherein the outlet port is configured to deliver the enriched population directly to the mammal.

[0123] Embodiment 18 is the device of any one of embodiments 1-17, wherein at least one of the one or more pumping elements is a displacement pump.

[0124] Embodiment 19 is the device of embodiment 18, wherein the displacement pump is a reciprocating pump. Embodiment 20 is the device of embodiment 19, wherein the reciprocating pump is a diaphragm pump.

[0125] Embodiment 21 is the device of embodiment 18, wherein the displacement pump is a rotary or continuous pump.

[0126] Embodiment 22 is the device of embodiment 21, wherein the displacement pump is a peristaltic pump.

[0127] Embodiment 23 is the device of any one of embodiments 1-22, wherein no component of any of the one or more pumping elements is located within the conduit.

[0128] Embodiment 24 is the device of any one of embodiments 1-23, wherein no component of any of the one or more pumping elements contacts the biological material or the dispersed form of the biological material when the biological material and the dispersed form of the biological material are within the conduit.

[0129] Embodiment 25 is the device of any one of embodiments 1-24, wherein the one or more dispersing elements are bottlenecks in the conduit.

[0130] Embodiment 26 is the device of embodiment 25, wherein the bottlenecks are compressed sections of the conduit.

[0131] Embodiment 27 is the device of embodiment 25, wherein the conduit comprises two or more bottlenecks separated by intervening segments of the conduit, and wherein the bottlenecks have a reduced maximum diameter as compared to the intervening segments.

[0132] Embodiment 28 is the device of any one of embodiments 25-27, wherein the conduit comprises 5 to 40 bottlenecks.

[0133] Embodiment 29 is the device of any one of embodiments 25-27, wherein the conduit comprises 20 to 30 bottlenecks.

[0134] Embodiment 30 is the device of embodiment 29, wherein the conduit is configured as a series of loops.

[0135] Embodiment 31 is the device of embodiment 30, wherein each loop comprises 4 to 6 bottlenecks.

[0136] Embodiment 32 is the device of embodiment 30 or embodiment 31, wherein each loop is configured to interact with a pumping element. Embodiment 33 is the device of any one of embodiments 1-32, wherein at least one of the one or more dispersing elements comprises one or more microstructures on an inner surface of the conduit.

[0137] Embodiment 34 is the device of any one of embodiments 1-33, wherein at least one of one or more separating elements is a filter.

[0138] Embodiment 35 is the device of embodiment 34, wherein the filter is a tangential flow filter.

[0139] Embodiment 36 is the device of embodiment 35, wherein the tangential flow filter is a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores.

[0140] Embodiment 37 is the device of any one of embodiments 1-36, wherein the one or more separating elements are filters in an axial arrangement.

[0141] Embodiment 38 is the device of any one of embodiments 1-36, wherein the one or more separating elements are filters in a radial arrangement.

[0142] Embodiment 39 is the device of any one of embodiments 1-36, wherein the one or more separating elements are filters in a parallel arrangement.

[0143] Embodiment 40 is the device of any one of embodiments 1-36, wherein the one or more separating elements are filters in a serial arrangement.

[0144] Embodiment 41 is the device of any one of embodiments 1-36, wherein the one or more separating elements are filters in a serial and parallel arrangement.

[0145] Embodiment 42 is the device of any one of embodiments 1-41, wherein the one or more separating elements comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger than about 80 to 100 pm if present in the dispersed form of the biological material from the enriched population.

[0146] Embodiment 43 is the device of embodiment 42, wherein the larger items comprise adipose cells.

[0147] Embodiment 44 is the device of any one of embodiments 1-43, wherein the one or more separating elements comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in the dispersed form of the biological material from the enriched population. Embodiment 45 is the device of embodiment 44, wherein the smaller items comprise fat globules, erythrocytes, or fat globules and erythrocytes.

[0148] Embodiment 46 is the device of any one of embodiments 1-45, wherein the device comprises one or more first holding chambers located along the conduit between the inlet port and a first of the one or more dispersing elements and configured to accumulate and hold the biological material.

[0149] Embodiment 47 is the device of any one of embodiments 1-46, wherein the device comprises one or more second holding chambers located along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to accumulate and hold the dispersed form of the biological material.

[0150] Embodiment 48 is the device of any one of embodiments 1-47, wherein the device comprises one or more third holding chambers located along the conduit between a first of the one or more separating elements and the outlet port and configured to accumulate and hold the enriched population.

[0151] Embodiment 49 is the device of any one of embodiments 1-48, wherein the device comprises one or more inflow ports fluidly connected to the conduit along the conduit between the inlet port and a first of the one or more dispersing elements and configured to add a fluid to the biological material.

[0152] Embodiment 50 is the device of embodiment 49, wherein the fluid is water or saline.

[0153] Embodiment 51 is the device of any one of embodiments 1-50, wherein the device comprises one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more dispersing elements and a first of the one or more separating elements and configured to add a fluid to the biological material.

[0154] Embodiment 52 is the device of embodiment 51, wherein the fluid is water or saline.

[0155] Embodiment 53 is the device of any one of embodiments 1-52, wherein the device comprises one or more inflow ports fluidly connected to the conduit along the conduit between a first of the one or more separating elements and the outlet port and configured to add a fluid to the enriched population.

[0156] Embodiment 54 is the device of embodiment 53, wherein the fluid is water or saline.

[0157] Embodiment 55 is the device of any one of embodiments 1-49, wherein the device lacks any inflow ports fluidly connected to the conduit along the conduit between the inlet port and the outlet port.

[0158] Embodiment 56 is the device of any one of embodiments 1-55, wherein the device comprises no more than one inflow port fluidly connected to the conduit along the conduit between the inlet port and the outlet port.

[0159] Embodiment 57 is the device of any one of embodiments 1-56, wherein the enriched population comprises about 150,000 cells / cm3.

[0160] Embodiment 58 is the device of any one of embodiments 1-57, wherein the enriched population of cells comprises at least 5-fold to 10-fold more cells per mg of starting biological material than the dispersed form of the biological material.

[0161] Embodiment 59 is a device for obtaining an enriched population of adipose- derived stem cells from biological material obtained from a mammal and returning the enriched population to the mammal, wherein the device comprises: (a) a conduit comprising an inlet port configured to receive the biological material from the mammal and an outlet port configured to deliver the enriched population to the mammal, (b) one or more pumping elements configured to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) one or more dispersing elements located along the conduit and configured to form a more dispersed form of the biological material from the biological material, wherein the dispersed form of the biological material comprises intact adipose-derived stem cells, and (d) one or more separating elements located along the conduit downstream of the one or more dispersing elements and configured to form the enriched population of adipose-derived stem cells from the dispersed form of the biological material, wherein the enriched population of adipose- derived stem cells comprises a greater number of adipose-derived stem cells per volume than the dispersed form of the biological material and comprises less non-cell material per volume than the dispersed form of the biological material.

[0162] Embodiment 60 is a method for preparing an enriched population of cells using a device of any one of embodiments 1-59, wherein the method comprises: (a) passing the biological material of the mammal through the inlet port and into the conduit, (b) actuating the one or more pumping elements to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) actuating the one or more dispersing elements to form the dispersed form of the biological material from the biological material, and (d) allowing the one or more separating elements to form the enriched population of cells from the dispersed form of the biological material.

[0163] Embodiment 61 is the method of embodiment 60, wherein the biological material is passed through the inlet port and into the conduit without purifying the biological material, without adding any preservatives to the biological material, and without diluting the biological material.

[0164] Embodiment 62 is the method of embodiment 60 or embodiment 61, comprising passing the biological material through the inlet port and into the conduit no more than about 5 minutes after the biological material was removed from the mammal.

[0165] Embodiment 63 is the method of any one of embodiments 60 to 62, wherein the method does not comprise centrifuging the biological material or the enriched population of cells.

[0166] Embodiment 64 is a method for delivering cells to a mammal using a device of any one of embodiments 1-59, wherein the method comprises: (a) passing the biological material of the mammal through the inlet port and into the conduit, (b) actuating the one or more pumping elements to move the biological material within the conduit in a direction from the inlet port to the outlet port, (c) actuating the one or more dispersing elements to form the dispersed form of the biological material from the biological material, (d) allowing the one or more separating elements to form the enriched population of cells from the dispersed form of the biological material, and (e) administering the enriched population of cells to the mammal. Embodiment 65 is the method of embodiment 64, wherein the biological material is passed through the inlet port and into the conduit without purifying the biological material, without adding any preservatives to the biological material, and without diluting the biological material.

[0167] Embodiment 66 is the method of embodiment 64 or embodiment 65, comprising passing the biological material through the inlet port and into the conduit no more than about 5 minutes after the biological material was removed from the mammal.

[0168] Embodiment 67 is the method of any one of embodiments 64 to 66, wherein the method does not comprise centrifuging the biological material or the enriched population of cells.

[0169] Embodiment 68 is the method of any one of embodiments 64 to 67, wherein the enriched population of cells is administered to the mammal without exposing the enriched population of cells to open air.

[0170] Embodiment 69 is the method of any one of embodiments 64 to 68, comprising administering the enriched population of cells to the mammal within about 5 to about 30 minutes after the biological material was obtained from the mammal.

[0171] Embodiment 70 is the method of any one of embodiments 64 to 69, wherein the method does not comprise culturing the biological materials or the enriched population of cells.

[0172] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0173] EXAMPLES Example 1 - Device for harvesting human fat and isolating adipose-derived stem cells therefrom

[0174] Biological material is completely conveyed within a closed system of tubes, filters, reservoirs, and ports and concentrated according to the desired component. This closed system is placed as a disposable item in a machine, which can be reused accordingly many times. The machine is housed in a rollable stainless steel enclosure that requires a footprint of about 0.4 x 0.4 m with a height of about 1.4 m. All surfaces are compatible with standard cleaning and disinfecting agents. The power supply is limited to a normal household power connection. In addition, a WLAN or LAN connection is available at the so-called “point-of-care” to enable gapless batch tracing of the sterile disposable set used.

[0175] The device is designed to contain:

[0176] • a power supply,

[0177] • a control system including a barcode and / or RFID scanner and a WLAN / Lan module,

[0178] • a drive technology (at least one peristaltic pump with one or more pump impellers, also called channels),

[0179] • one or more fastening options for the patient-specific disposable sets,

[0180] • one or more squeeze points for the flexible tubing of the disposable set, and

[0181] • one or more additional holders for medical bags and bottles.

[0182] At this device, after a capture of the barcode and / or RFID information of the disposable set and release of the same through a cloud-based database, the disposable set is released for use, and the peristaltic pump(s) are released and preset according to the disposable set.

[0183] The complete set, including inlet and outlet ports, a flexible tube as conduit, at least two separating elements (tangential flow filters, such as hollow fiber filters), and reservoirs, is then attached to the machine. In addition (e.g., if needed for space reasons), the tube is laid on the peristaltic pump in several loops around the individual pump impellers (channels) and simultaneously fixed in numerous receptacles. In these receptacles, the tubing is squeezed (e.g., strongly squeezed) by mechanical tensioning so that the round cross-section is compressed into a narrow, elongated channel cross-section.

[0184] When biological material is pumped by the peristaltic pump in the flexible tube at a few bar, an increased shear stress occurs in these squeezing points, which in turn gently disperses the biological material. To facilitate handling by the operator, several pinch points are locked simultaneously, each with a movable element. The correct setting of the numerous squeezing points is sensor-monitored in the sense of error-proofing. Once the complete set is inserted or fastened, the additional materials are connected to their respective ports. These can be, for example, additional water, saline solution, but also other biological and / or medical materials.

[0185] Once the disposable set is connected to a material dispenser (e.g., the outlet of a liposuction machine, a bag containing body fat, etc.) and a material recipient (e.g., a receiving bag, a syringe, or a patient, etc.), the process for enriching biological material is initiated. Biological material is continuously and promptly conveyed, dispersed, and concentrated depending on the desired component (e.g., for enrichment of adipose mesenchymal stem cells) in a completely closed, sterile process directly on site and / or close to the patient.

[0186] If biological material is collected directly from a mammal (e.g., a human or a horse) on site, the entire process, including a delivery of the enriched biological material back to the donor, can be designed to take no more than about 5 to 30 minutes.

[0187] For hygienic reasons, a new, completely prefabricated set is used for each treatment in order to reduce any contamination or cross-contamination.

[0188] Example 2 - Device for harvesting human fat and isolating adipose-derived stem cells therefrom

[0189] Biological material is completely conveyed within a closed system of tubes, filters, reservoirs, and ports and concentrated according to the desired component. This closed system is placed as a disposable item in a machine, which can be reused accordingly many times.

[0190] The machine is housed in a rollable stainless steel enclosure that requires a footprint of about 0.4 x 0.4 m with a height of about 1.4 m. All surfaces are compatible with standard cleaning and disinfecting agents. The power supply is limited to a normal household power connection. In addition, a WLAN or LAN connection is available at the so-called “point-of-care” to enable gapless batch tracing of the sterile disposable set used.

[0191] The device is designed to contain:

[0192] • a power supply, a control system including a barcode and / or RFID scanner and a WLAN / Lan module,

[0193] • a drive technology (at least one peristaltic pump with one or more pump impellers, also called channels),

[0194] • one or more fastening options for the patient-specific disposable sets,

[0195] • one or more squeeze points for the flexible tubing of the disposable set, and

[0196] • one or more additional holders for medical bags and bottles.

[0197] At this device, after a capture of the barcode and / or RFID information of the disposable set and release of the same through a cloud-based database, the disposable set is released for use, and the peristaltic pump(s) are released and preset according to the disposable set.

[0198] The complete set, including inlet and outlet ports, a flexible tube as conduit, at least two separating stages (either integrated in a module or in separate filter housings), and, if needed reservoirs, is then attached to the machine. The biological material flows through at least two filter stages (e.g., a flow filtration stage and a tangential flow filtration stage) one after the other. In addition (e.g., if needed for space reasons), the tube is laid on the peristaltic pump in several loops around the individual pump impellers (channels) and simultaneously fixed in numerous receptacles. In these receptacles, the tubing is squeezed by mechanical tensioning so that the round cross-section is compressed into a narrow, elongated channel cross-section.

[0199] When biological material is pumped by the peristaltic pump in the flexible tube at a few bar, an increased shear stress occurs in these squeezing points, which in turn gently disperses the biological material. To facilitate handling by the operator, several pinch points are locked simultaneously, each with a movable element (e.g., a spring-mounted element that contacts an external surface of the flexible tube distal to the pump impeller(s), and / or a radially movable roller element that contacts an external surface of the flexible tube proximal to the pump impeller(s)).

[0200] Once the complete set is inserted or fastened, the additional materials are connected to their respective ports. These can be, for example, additional water, saline solution, but also other biological and / or medical materials. Once the disposable set is connected to a material dispenser (e.g., the outlet of a liposuction machine, a bag containing body fat, etc.) and a material recipient (e.g., a receiving bag, a syringe, or a patient, etc.), the process for enriching biological material is initiated. Biological material is continuously and promptly conveyed, dispersed, and concentrated depending on the desired component (e.g., for enrichment of adipose mesenchymal stem cells) in a completely closed, sterile process directly on site and / or close to the patient.

[0201] If biological material is collected directly from a mammal (e.g., a human or a horse) on site, the entire process, including a delivery of the enriched biological material back to the donor, can be designed to take no more than about 5 to 30 minutes.

[0202] For hygienic reasons, a new, completely prefabricated set is used for each treatment in order to reduce any contamination or cross-contamination.

[0203] Example 3 - Treatment of horses

[0204] A horse in need of treatment (e.g., a horse having tendon damage) is treated with a device provided herein in order to promote tendon regeneration. A veterinarian brings a sterilely packaged device provided herein, along with syringes, local anesthetics, and disinfectants, to the location of the horse to be treated (e.g., a veterinary clinic, a stable, or even an outdoor paddock or corral). The veterinarian cleans and disinfects the horse’s skin before beginning removal of biological material from the animal using a device provided herein (e.g., as described in Example 1 or Example 2). The biological material only comes into contact with originally sterile-packed elements from the time of removal until the concentrated biological material is reintroduced into the horse.

[0205] OTHER EMBODIMENTS

[0206] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A device for obtaining an enriched population of cells from biological material obtained from a mammal, wherein said device comprises:(a) a conduit comprising an inlet port configured to receive said biological material from said mammal and an outlet port configured to output said enriched population from said conduit,(b) one or more pumping elements configured to move said biological material within said conduit in a direction from said inlet port to said outlet port,(c) one or more dispersing elements located along said conduit and configured to form a more dispersed form of said biological material from said biological material, wherein said dispersed form of said biological material comprises intact cells, and(d) one or more separating elements located along said conduit downstream of said one or more dispersing elements and configured to form said enriched population of cells from said dispersed form of said biological material, wherein said enriched population of cells comprises a greater number of cells per volume than said dispersed form of said biological material and comprises less non-cell material per volume than said dispersed form of said biological material.

2. The device of claim 1, wherein said enriched population of cells is an enriched population of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells.

3. The device of claim 1, wherein said cells are adipose-derived stem cells.

4. The device of claim 1, wherein said biological material comprises fat tissue.

5. The device of claim 1, wherein said mammal is a human.

6. The device of claim 1, wherein said mammal is a horse.

7. The device of claim 1, wherein said conduit comprises flexible tubing.

8. The device of claim 1, wherein said conduit is from about 20 cm to about 10 m in length.

9. The device of claim 1, wherein said conduit comprises at least one section having an inner diameter of about 1 mm to 5 mm.

10. The device of claim 1, wherein a maximum inner diameter along any point of said conduit is no greater than 20 mm.

11. The device of claim 1, wherein a minimum inner diameter along any point of said conduit is no less than 0.1 mm.

12. The device of claim 1, wherein said conduit comprises stainless steel, ceramic, glass, a thermosetting plastic, a thermoplastic polymer, an elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, a thermoplastic elastomer, a urethane polyethersulfone, polysulfone, polyacrylnitrile, polycarbonate, rubber, or any combination thereof.

13. The device of claim 1, wherein said inlet port is configured to attach to an outlet of a liposuction device.

14. The device of claim 13, further comprising an adaptor configured to attach said inlet port to said liposuction device.

15. The device of claim 1, wherein said inlet port is configured to collect said biological material directly from said mammal.

16. The device of claim 1, wherein said outlet port is configured to attach to an inlet of an administering device.

17. The device of claim 1, wherein said outlet port is configured to deliver said enriched population directly to said mammal.

18. The device of claim 1, wherein at least one of said one or more pumping elements is a displacement pump.

19. The device of claim 18, wherein said displacement pump is a reciprocating pump.

20. The device of claim 19, wherein said reciprocating pump is a diaphragm pump.

21. The device of claim 18, wherein said displacement pump is a rotary or continuous pump.

22. The device of claim 21, wherein said displacement pump is a peristaltic pump.

23. The device of claim 1, wherein no component of any of said one or more pumping elements is located within said conduit.

24. The device of claim 1, wherein no component of any of said one or more pumping elements contacts said biological material or said dispersed form of said biological material when said biological material and said dispersed form of said biological material are within said conduit.

25. The device of claim 1, wherein said one or more dispersing elements are bottlenecks in said conduit.

26. The device of claim 25, wherein said bottlenecks are compressed sections of said conduit.

27. The device of claim 25, wherein said conduit comprises two or more bottlenecks separated by intervening segments of said conduit, and wherein said bottlenecks have a reduced maximum diameter as compared to said intervening segments.

28. The device of claim 25, wherein said conduit comprises 5 to 40 bottlenecks.

29. The device of claim 25, wherein said conduit comprises 20 to 30 bottlenecks.

30. The device of claim 29, wherein said conduit is configured as a series of loops.

31. The device of claim 30, wherein each loop comprises 4 to 6 bottlenecks.

32. The device of claim 30, wherein each loop is configured to interact with a pumping element.

33. The device of claim 1, wherein at least one of said one or more dispersing elements comprises one or more microstructures on an inner surface of said conduit.

34. The device of claim 1, wherein at least one of one or more separating elements is a filter.

35. The device of claim 34, wherein said filter is a tangential flow filter.

36. The device of claim 35, wherein said tangential flow filter is a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures that are separated from one another to define pores.

37. The device of claim 1, wherein said one or more separating elements are filters in an axial arrangement.

38. The device of claim 1, wherein said one or more separating elements are filters in a radial arrangement.

39. The device of claim 1, wherein said one or more separating elements are filters in a parallel arrangement.

40. The device of claim 1, wherein said one or more separating elements are filters in a serial arrangement.

41. The device of claim 1, wherein said one or more separating elements are filters in a serial and parallel arrangement.

42. The device of claim 1, wherein said one or more separating elements comprise at least one filter having a pore size of about 80 to 100 pm to remove items larger thanabout 80 to 100 pm if present in said dispersed form of said biological material from said enriched population.

43. The device of claim 42, wherein said larger items comprise adipose cells.

44. The device of claim 1, wherein said one or more separating elements comprise at least one filter having a pore size of about 10 pm to remove items smaller than about 10 pm if present in said dispersed form of said biological material from said enriched population.

45. The device of claim 44, wherein said smaller items comprise fat globules, erythrocytes, or fat globules and erythrocytes.

46. The device of claim 1, wherein said device comprises one or more first holding chambers located along said conduit between said inlet port and a first of said one or more dispersing elements and configured to accumulate and hold said biological material.

47. The device of claim 1, wherein said device comprises one or more second holding chambers located along said conduit between a first of said one or more dispersing elements and a first of said one or more separating elements and configured to accumulate and hold said dispersed form of said biological material.

48. The device of claim 1, wherein said device comprises one or more third holding chambers located along said conduit between a first of said one or more separating elements and said outlet port and configured to accumulate and hold said enriched population.

49. The device of claim 1, wherein said device comprises one or more inflow ports fluidly connected to said conduit along said conduit between said inlet port and a first of said one or more dispersing elements and configured to add a fluid to said biological material.

50. The device of claim 49, wherein said fluid is water or saline.

51. The device of claim 1, wherein said device comprises one or more inflow ports fluidly connected to said conduit along said conduit between a first of said one or more dispersing elements and a first of said one or more separating elements and configured to add a fluid to said biological material.

52. The device of claim 51, wherein said fluid is water or saline.

53. The device of claim 1, wherein said device comprises one or more inflow ports fluidly connected to said conduit along said conduit between a first of said one or more separating elements and said outlet port and configured to add a fluid to said enriched population.

54. The device of claim 53, wherein said fluid is water or saline.

55. The device of claim 1, wherein said device lacks any inflow ports fluidly connected to said conduit along said conduit between said inlet port and said outlet port.

56. The device of claim 1, wherein said device comprises no more than one inflow port fluidly connected to said conduit along said conduit between said inlet port and said outlet port.

57. The device of claim 1, wherein said enriched population comprises about 150,000 cells / cm3.

58. The device of claim 1, wherein said enriched population of cells comprises at least 5-fold to 10-fold more cells per mg of starting biological material than said dispersed form of said biological material.

59. A device for obtaining an enriched population of adipose-derived stem cells from biological material obtained from a mammal and returning said enriched population to said mammal, wherein said device comprises:(a) a conduit comprising an inlet port configured to receive said biological material from said mammal and an outlet port configured to deliver said enriched population to said mammal,(b) one or more pumping elements configured to move said biological material within said conduit in a direction from said inlet port to said outlet port,(c) one or more dispersing elements located along said conduit and configured to form a more dispersed form of said biological material from said biological material, wherein said dispersed form of said biological material comprises intact adipose-derived stem cells, and(d) one or more separating elements located along said conduit downstream of said one or more dispersing elements and configured to form said enriched population of adipose-derived stem cells from said dispersed form of said biological material, wherein said enriched population of adipose-derived stem cells comprises a greater number of adipose-derived stem cells per volume than said dispersed form of said biological material and comprises less non-cell material per volume than said dispersed form of said biological material.

60. A method for preparing an enriched population of cells using the device of claim 1, wherein said method comprises:(a) passing said biological material of said mammal through said inlet port and into said conduit,(b) actuating said one or more pumping elements to move said biological material within said conduit in a direction from said inlet port to said outlet port,(c) actuating said one or more dispersing elements to form said dispersed form of said biological material from said biological material, and(d) allowing said one or more separating elements to form said enriched population of cells from said dispersed form of said biological material.

61. The method of claim 60, wherein said biological material is passed through said inlet port and into said conduit without purifying said biological material, without addingany preservatives to said biological material, and without diluting said biological material.

62. The method of claim 60, comprising passing said biological material through said inlet port and into said conduit no more than about 5 minutes after said biological material was removed from said mammal.

63. The method of claim 60, wherein said method does not comprise centrifuging said biological material or said enriched population of cells.

64. A method for delivering cells to a mammal using the device of claim 1, wherein said method comprises:(a) passing said biological material of said mammal through said inlet port and into said conduit,(b) actuating said one or more pumping elements to move said biological material within said conduit in a direction from said inlet port to said outlet port,(c) actuating said one or more dispersing elements to form said dispersed form of said biological material from said biological material,(d) allowing said one or more separating elements to form said enriched population of cells from said dispersed form of said biological material, and(e) administering said enriched population of cells to said mammal.

65. The method of claim 64, wherein said biological material is passed through said inlet port and into said conduit without purifying said biological material, without adding any preservatives to said biological material, and without diluting said biological material.

66. The method of claim 64, comprising passing said biological material through said inlet port and into said conduit no more than about 5 minutes after said biological material was removed from said mammal.

67. The method of claim 64, wherein said method does not comprise centrifuging said biological material or said enriched population of cells.

68. The method of claim 64, wherein said enriched population of cells is administered to said mammal without exposing said enriched population of cells to open air.

69. The method of claim 64, comprising administering said enriched population of cells to said mammal within about 5 to about 30 minutes after said biological material was obtained from said mammal.

70. The method of claim 64, wherein said method does not comprise culturing said biological materials or said enriched population of cells.

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