Apparatus and system for receiving biological material and providing an enriched component of the biological material

Abstract

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

Description

Cross-reference to related applications

[0001] This application claims priority to U.S. Provisional Application Serial No. 63 / 527,149, filed July 17, 2023. The disclosure of the earlier application is considered part of the disclosure of this application (and is incorporated herein by reference). Technical Field

[0002] This document relates to devices and systems (e.g., bedside devices and systems) designed to receive biological material (e.g., adipose tissue) from a mammal (e.g., a human or horse) and to process the biological material to provide an end product comprising one or more enriched components of the received biological material, which may optionally be reintroduced into the same mammal. For example, this document provides bedside devices designed to collect adipose tissue from a mammal (e.g., a human) in a clinical setting and process the adipose tissue to provide adipose-derived stem cells as an enriched end product of the collected adipose tissue, which may optionally be reintroduced into the same mammal in a clinical setting. This document also provides methods for obtaining biological material (e.g., adipose tissue) from a mammal (e.g., a human) and processing the biological material to provide an end product comprising one or more enriched components of the received biological material, which may optionally be reintroduced into the same mammal. Background Technology

[0003] Both adipose-derived stem cells (also known as adipose-derived regenerative cells, adipose-derived adult stem cells, and adipose-derived mesenchymal stem cells) and stromal vascular fractions have attracted interest in therapeutic applications within the fields 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 (Lee et al., 114:108765, 2019) demonstrated the general potential of adipose-derived stem cells in regenerative medicine, particularly in areas such as bone / cartilage repair, muscle regeneration, and osteoarthritis treatment (Lee et al., 114:108765, 2019). 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 potential for treatment of many other conditions. Interest in these pluripotent stem cells and stromal vascular components is based on the possibility of obtaining large quantities of cells and materials through minimally invasive surgery. Other forms of stem cell therapy, such as bone marrow stem cell therapy and umbilical cord stem cell therapy, may have higher risks associated with collection, potentially high morbidity, difficulty in obtaining sufficient cell quantities, and may raise thorny ethical and regulatory issues. On the other hand, adipose-derived stem cells can be obtained not only from solid fat but also from fat aspirates. The lack of immune response, the ability of cells to self-renew, their similarity to the natural healing process, and the ability of cells to develop into desired tissues are of particular interest. The ability of adipose-derived stem cells to differentiate into different tissue-specific progenitor cells, migrate to damaged sites, and function through autocrine and paracrine pathways suggests that these cells may meet the criteria for successful cell therapy. Summary of the Invention

[0004] The lack of standardized protocols for cell management in laboratories and operating rooms, along with the non-sterile, complex, and slow process of isolating adipose-derived stem cells, limits their widespread use by balancing their real-life benefits with patient safety and security.

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

[0006] As described herein, a harvesting system can be designed to harvest biological material (e.g., adipose tissue) from a mammal (e.g., a human) and process the biological material to form a viscous fluid comprising intact cells. The system can also be designed to process the viscous fluid to form an end product enriched with 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, the closed-loop system described herein can be used to harvest biological material (e.g., adipose tissue) from a human, process the harvested biological material to obtain an end product enriched with adipose-derived stem cells, and administer the obtained end product directly back into the human body.

[0007] Generally speaking, one aspect of this document provides an apparatus for obtaining a rich cluster of cells from biological material obtained from a mammal. The apparatus comprises (or is substantially composed of, or consists of): (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the rich cluster 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 positioned 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 includes intact cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the rich cluster of cells from the dispersed form of the biological material, wherein the rich cluster of cells per volume comprises a greater number of cells than the dispersed form of the biological material, and comprises less non-cellular material per volume than the dispersed form of the biological material. The enriched cell population may be an enriched cluster of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells may be adipose-derived stem cells. The biomaterial may include adipose tissue. The mammal may be human. The mammal may be a horse. The catheter may include a flexible tube. The length of the catheter may be from about 20 cm to about 10 m. The catheter may include at least one segment with an inner diameter of about 1 mm to 5 mm. In some cases, the maximum inner diameter at any point along the catheter may not be greater than 20 mm. In some cases, the minimum inner diameter at any point along the catheter may not be less than 0.1 mm. The catheter may include stainless steel, ceramic, glass, thermosetting plastic, thermoplastic polymer, elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomer, urethane polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber, or any combination thereof. The inlet port may be configured to attach to the outlet of a liposuction device. The device may include an adapter configured to attach the inlet port to the liposuction device. The inlet port may be configured to collect biomaterial directly from the mammal. The outlet port may be configured to attach to the inlet of an application device. The outlet port can be configured to deliver the enriched cluster directly to the mammal. At least one of the one or more pumping elements can be a positive displacement pump. A positive displacement pump can be a reciprocating pump. A reciprocating pump can be a diaphragm pump. A positive displacement pump can be a rotary pump or a continuous pump. A positive displacement pump can be a peristaltic pump. In some cases, components of any one of the one or more pumping elements are not located within the conduit. In some cases, when the biological material and its dispersion are located within the conduit, components of any one of the one or more pumping elements do not contact the biological material or its dispersion. One or more dispersing elements can be bottlenecks in the conduit.The bottleneck may be a compression section of the conduit. The conduit may include two or more bottlenecks separated by a middle section, wherein the bottlenecks may have a reduced maximum diameter compared to the middle section. The conduit may include 5 to 40 bottlenecks. The conduit may include 20 to 30 bottlenecks. The conduit may be configured as a series of loops. Each loop may include 4 to 6 bottlenecks. Each loop may be configured to interact with a pumping element. At least one of the one or more dispersing elements may include one or more microstructures located on the inner surface of the conduit. At least one of the one or more separating elements may be a filter. The filter may be a tangential flow filter. A tangential flow filter may be a hollow fiber filter, a diaphragm filter, or a filter having multiple integrated structures spaced apart to define pores. One or more separating elements may be filters arranged axially. One or more separating elements may be filters arranged radially. One or more separating elements may be filters arranged in parallel. One or more separating elements may be filters arranged in series. One or more separating elements may be filters arranged in both series and parallel. One or more separation elements may include at least one filter with a pore size of about 80 to 100 μm to remove material from the rich cluster if material larger than about 80 to 100 μm is present in the dispersed form of the biological material. Larger material may include fat cells. One or more separation elements may include at least one filter with a pore size of about 10 μm to remove material from the rich cluster if material smaller than about 10 μm is present in the dispersed form of the biological material. Smaller material includes fat globules, erythrocytes, or both fat globules and erythrocytes. The device may include one or more first holding chambers located along a conduit between an inlet port and a first dispersing element of one or more dispersion elements, and configured to accumulate and retain the biological material. The device may include one or more second holding chambers located along a conduit between a first dispersing element of one or more dispersion elements and a first separating element of one or more separation elements, and configured to accumulate and retain the dispersed form of the biological material. The device may include one or more third holding chambers located along a conduit between a first separating element of one or more separation elements and an outlet port, and configured to accumulate and retain the rich cluster. The device may include one or more inflow ports that are fluidly connected to a conduit along a conduit located between an inlet port and a first dispersing element of one or more dispersing elements, and is configured to add fluid to biological material. The fluid may be water or brine.The device may include one or more inflow ports fluidly connected to a conduit along a conduit located between a first separating element and an outlet port in one or more separating elements, and is configured to add fluid to the enriched cluster. The fluid may be water or brine. The device may lack any inflow ports fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The device may include no more than one inflow port fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The enriched cluster may include approximately 150,000 cells / cm². 3 Cell-rich clusters may include at least 5 to 10 times more cells per mg of starting biomaterial than in dispersed forms of biomaterial.

[0008] On the other hand, this document provides an apparatus for obtaining a rich cluster of adipose-derived stem cells from biological material obtained from a mammal and returning the rich cluster to the mammal. The apparatus comprises (or is substantially composed of, or consists of): (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to deliver the rich cluster 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 positioned 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 includes intact adipose-derived stem cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the rich cluster of adipose-derived stem cells from the dispersed form of the biological material, wherein the rich cluster of adipose-derived stem cells per volume includes a greater number of adipose-derived stem cells than the dispersed form of the biological material, and per volume includes less non-cellular material than the dispersed form of the biological material. The cell-rich clusters may be rich clusters of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells may be adipose-derived stem cells. The biomaterial may include adipose tissue. The mammal may be human. The catheter may include a flexible tube. The length of the catheter may be from about 20 cm to about 10 m. The catheter may include at least one segment with an inner diameter of about 1 mm to 5 mm. In some cases, the maximum inner diameter at any point along the catheter may not be greater than 20 mm. In some cases, the minimum inner diameter at any point along the catheter may not be less than 0.1 mm. The catheter may include stainless steel, ceramic, glass, thermosetting plastics, thermoplastic polymers, elastomers, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomers, urethane polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber, or any combination thereof. The inlet port may be configured to attach to the outlet of a liposuction device. The device may include an adapter configured to attach the inlet port to the liposuction device. The inlet port may be configured to collect biomaterial directly from the mammal. The outlet port may be configured to attach to the inlet of an application device. The outlet port can be configured to deliver the rich cluster directly to the mammal. At least one of the one or more pumping elements can be a positive displacement pump. The positive displacement pump can be a reciprocating pump. The reciprocating pump can be a diaphragm pump. The positive displacement pump can be a rotary pump or a continuous pump. The positive displacement pump can be a peristaltic pump. In some cases, components of any one of the one or more pumping elements are not located within the conduit.In some cases, when the biomaterial and its dispersion are located within a catheter, components of any one or more pumping elements do not contact the biomaterial or its dispersion. One or more dispersing elements may be bottlenecks in the catheter. A bottleneck may be a compression section of the catheter. The catheter may include two or more bottlenecks separated by a mid-section, wherein the bottlenecks may have a reduced maximum diameter compared to the mid-section. The catheter may include 5 to 40 bottlenecks. The catheter may include 20 to 30 bottlenecks. The catheter may be configured as a series of loops. Each loop may include 4 to 6 bottlenecks. Each loop may be configured to interact with a pumping element. At least one of the one or more dispersing elements may include one or more microstructures located on the inner surface of the catheter. At least one of the one or more separating elements may be a filter. The filter may be a tangential flow filter. A tangential flow filter may be a hollow fiber filter, a diaphragm filter, or a filter having multiple integrated structures spaced apart to define pores. One or more separating elements may be filters arranged axially. One or more separating elements may be filters arranged radially. One or more separating elements may be filters arranged in parallel. One or more separating elements may be filters arranged in series. One or more separating elements may be filters arranged in both series and parallel. One or more separating elements may include at least one filter with a pore size of about 80 to 100 μm to remove material from the rich clusters if material larger than about 80 to 100 μm is present in the dispersed form of the biological material. Larger material may include fat cells. One or more separating elements may include at least one filter with a pore size of about 10 μm to remove material from the rich clusters if material smaller than about 10 μm is present in the dispersed form of the biological material. Smaller material includes fat globules, erythrocytes, or both fat globules and erythrocytes. The device may include one or more first holding chambers located along a conduit between an inlet port and a first dispersing element of one or more dispersing elements, and configured to accumulate and retain the biological material. The device may include one or more second holding chambers located along a conduit between a first dispersing element of one or more dispersing elements and a first separating element of one or more separating elements, and configured to accumulate and retain the dispersed form of the biological material. The device includes one or more third retaining chambers located along a conduit between a first separating element and an outlet port of one or more separating elements, and is configured to accumulate and retain the rich cluster. The device may include one or more inflow ports fluidly connected to the conduit along a conduit located between an inlet port and a first dispersing element of one or more dispersing elements, and is configured to add fluid to the biomaterial. The fluid may be water or brine.The device may include one or more inflow ports fluidly connected to a conduit along a conduit between a first dispersing element and a first separating element in one or more dispersing elements, and is configured to add fluid to the biological material. The fluid may be water or saline. The device may also include one or more inflow ports fluidly connected to a conduit along a conduit between a first separating element and an outlet port in one or more dispersing elements, and is configured to add fluid to a rich cluster. The fluid may be water or saline. The device may lack any inflow ports fluidly connected to the conduit along a conduit between an inlet port and an outlet port. The device may include no more than one inflow port fluidly connected to the conduit along a conduit between an inlet port and an outlet port. The rich cluster may include approximately 150,000 cells / cm². 3 Cell-rich clusters may include at least 5 to 10 times more cells per mg of starting biomaterial than in dispersed forms of biomaterial.

[0009] On the other hand, this document provides a method for preparing a rich cluster of cells using an apparatus for obtaining a rich cluster of cells from biological material obtained from a mammal. The apparatus may include (or may consist substantially of, or may consist of): (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the rich cluster 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 positioned 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 includes intact cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the rich cluster of cells from the dispersed form of the biological material, wherein the rich cluster of cells per volume includes a greater number of cells than the dispersed form of the biological material, and a less non-cellular material per volume than the dispersed form of the biological material. The cell-rich clusters may be rich clusters of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells may be adipose-derived stem cells. The biomaterial may include adipose tissue. The mammal may be human. The catheter may include a flexible tube. The length of the catheter may be from about 20 cm to about 10 m. The catheter may include at least one segment with an inner diameter of about 1 mm to 5 mm. In some cases, the maximum inner diameter at any point along the catheter may not exceed 20 mm. In some cases, the minimum inner diameter at any point along the catheter may not be less than 0.1 mm. The catheter may include stainless steel, ceramic, glass, thermosetting plastics, thermoplastic polymers, elastomers, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomers, urethane polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber, or any combination thereof. The inlet port may be configured to attach to the outlet of a liposuction device. The device may include an adapter configured to attach the inlet port to the liposuction device. The inlet port may be configured to collect biomaterial directly from the mammal. The outlet port may be configured to attach to the inlet of an application device. The outlet port can be configured to deliver the enriched cluster directly to the mammal. At least one of the one or more pumping elements can be a positive displacement pump. A positive displacement pump can be a reciprocating pump. A reciprocating pump can be a diaphragm pump. A positive displacement pump can be a rotary pump or a continuous pump. A positive displacement pump can be a peristaltic pump. In some cases, components of any one of the one or more pumping elements are not located within the conduit. In some cases, when the biological material and its dispersion are located within the conduit, components of any one of the one or more pumping elements do not contact the biological material or its dispersion. One or more dispersing elements can be bottlenecks in the conduit.The bottleneck may be a compression section of the conduit. The conduit may include two or more bottlenecks separated by a middle section, wherein the bottlenecks may have a reduced maximum diameter compared to the middle section. The conduit may include 5 to 40 bottlenecks. The conduit may include 20 to 30 bottlenecks. The conduit may be configured as a series of loops. Each loop may include 4 to 6 bottlenecks. Each loop may be configured to interact with a pumping element. At least one of the one or more dispersing elements may include one or more microstructures located on the inner surface of the conduit. At least one of the one or more separating elements may be a filter. The filter may be a tangential flow filter. A tangential flow filter may be a hollow fiber filter, a diaphragm filter, or a filter having multiple integrated structures spaced apart to define pores. One or more separating elements may be filters arranged axially. One or more separating elements may be filters arranged radially. One or more separating elements may be filters arranged in parallel. One or more separating elements may be filters arranged in series. One or more separating elements may be filters arranged in both series and parallel. One or more separation elements may include at least one filter with a pore size of about 80 to 100 μm to remove material from the rich cluster if material larger than about 80 to 100 μm is present in the dispersed form of the biological material. Larger material may include fat cells. One or more separation elements may include at least one filter with a pore size of about 10 μm to remove material from the rich cluster if material smaller than about 10 μm is present in the dispersed form of the biological material. Smaller material includes fat globules, erythrocytes, or both fat globules and erythrocytes. The device may include one or more first holding chambers located along a conduit between an inlet port and a first dispersing element of one or more dispersion elements, and configured to accumulate and retain the biological material. The device may include one or more second holding chambers located along a conduit between a first dispersing element of one or more dispersion elements and a first separating element of one or more separation elements, and configured to accumulate and retain the dispersed form of the biological material. The device may include one or more third holding chambers located along a conduit between a first separating element of one or more separation elements and an outlet port, and configured to accumulate and retain the rich cluster. The device may include one or more inflow ports that are fluidly connected to a conduit along a conduit located between an inlet port and a first dispersing element of one or more dispersing elements, and is configured to add fluid to biological material. The fluid may be water or brine.The device may include one or more inflow ports fluidly connected to a conduit along a conduit located between a first separating element and an outlet port in one or more separating elements, and is configured to add fluid to the enriched cluster. The fluid may be water or brine. The device may lack any inflow ports fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The device may include no more than one inflow port fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The enriched cluster may include approximately 150,000 cells / cm². 3 The cell-rich clusters may comprise at least 5 to 10 times more cells / mg of starting biological material than the dispersed form of the biological material. The method comprises (or substantially comprises, or consists of): (a) passing the mammalian biological material through an inlet port and into a conduit; (b) actuating 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 one or more dispersing elements to form a dispersed form of the biological material from the biological material; and (d) allowing one or more separating elements to form cell-rich clusters from the dispersed form of the biological material. The biological material may be passed through the inlet port and into the conduit without purification of the biological material, without adding any preservatives to the biological material, and without dilution of the biological material. The method may include passing the biological material through the inlet port and into the conduit no more than about 5 minutes after removal of the biological material from the mammal. The method may be one that does not involve centrifugation of the biological material or the cell-rich clusters.

[0010] On the other hand, this document provides a method for delivering cells to a mammal using a device. The device may include (or may consist substantially of, or may consist of): (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the rich cluster 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 positioned 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 includes intact cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the rich cluster of cells from the dispersed form of the biological material, wherein the rich cluster of cells per volume includes a greater number of cells than the dispersed form of the biological material, and less non-cellular material per volume than the dispersed form of the biological material. The rich cluster of cells may be a rich cluster of adipose-derived stem cells, mesenchymal stem cells, or hematopoietic stem cells. The cells may be adipose-derived stem cells. The biomaterial may include adipose tissue. The mammal may be human. The catheter may include a flexible tube. The length of the catheter may be from about 20 cm to about 10 m. The catheter may include at least one segment with an inner diameter of about 1 mm to 5 mm. In some cases, the maximum inner diameter at any point along the catheter may not be greater than 20 mm. In some cases, the minimum inner diameter at any point along the catheter may not be less than 0.1 mm. The catheter may include stainless steel, ceramic, glass, thermosetting plastic, thermoplastic polymer, elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomer, urethane polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber, or any combination thereof. The inlet port may be configured to attach to the outlet of the liposuction device. The device may include an adapter configured to attach the inlet port to the liposuction device. The inlet port may be configured to collect biomaterial directly from the mammal. The outlet port may be configured to attach to the inlet of the application device. The outlet port may be configured to deliver the biomaterial directly to the mammal. At least one of the one or more pumping elements may be a positive displacement pump. Positive displacement pumps can be reciprocating pumps. Reciprocating pumps can be diaphragm pumps. Positive displacement pumps can be rotary pumps or continuous pumps. Positive displacement pumps can be peristaltic pumps. In some cases, components of any one or more pumping elements are not located within the conduit. In some cases, when the biological material and its dispersion are located within the conduit, components of any one or more pumping elements do not contact the biological material or its dispersion. One or more dispersing elements can be bottlenecks in the conduit. Bottlenecks can be compression sections of the conduit.The conduit may include two or more bottleneck sections separated by a middle section of the conduit, wherein the bottleneck sections may have a reduced maximum diameter compared to the middle section. The conduit may include 5 to 40 bottleneck sections. The conduit may include 20 to 30 bottleneck sections. The conduit may be configured as a series of loops. Each loop may include 4 to 6 bottleneck sections. Each loop may be configured to interact with a pumping element. At least one of the one or more dispersing elements may include one or more microstructures located on the inner surface of the conduit. At least one of the one or more separating elements may be a filter. The filter may be a tangential flow filter. A tangential flow filter may be a hollow fiber filter, a diaphragm filter, or a filter having multiple integrated structures spaced apart to define pores. One or more separating elements may be filters arranged axially. One or more separating elements may be filters arranged radially. One or more separating elements may be filters arranged in parallel. One or more separating elements may be filters arranged in series. One or more separating elements may be filters arranged in both series and parallel. One or more separation elements may include at least one filter with a pore size of about 80 to 100 μm to remove material from the rich cluster if material larger than about 80 to 100 μm is present in the dispersed form of the biological material. Larger material may include fat cells. One or more separation elements may include at least one filter with a pore size of about 10 μm to remove material from the rich cluster if material smaller than about 10 μm is present in the dispersed form of the biological material. Smaller material includes fat globules, erythrocytes, or both fat globules and erythrocytes. The device may include one or more first holding chambers located along a conduit between an inlet port and a first dispersing element of one or more dispersion elements, and configured to accumulate and retain the biological material. The device may include one or more second holding chambers located along a conduit between a first dispersing element of one or more dispersion elements and a first separating element of one or more separation elements, and configured to accumulate and retain the dispersed form of the biological material. The device may include one or more third holding chambers located along a conduit between a first separating element of one or more separation elements and an outlet port, and configured to accumulate and retain the rich cluster. The device may include one or more inflow ports that are fluidly connected to a conduit along a conduit located between an inlet port and a first dispersing element of one or more dispersing elements, and is configured to add fluid to biological material. The fluid may be water or brine.The device may include one or more inflow ports fluidly connected to a conduit along a conduit located between a first separating element and an outlet port in one or more separating elements, and is configured to add fluid to the enriched cluster. The fluid may be water or brine. The device may lack any inflow ports fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The device may include no more than one inflow port fluidly connected to the conduit along a conduit located between the inlet and outlet ports. The enriched cluster may include approximately 150,000 cells / cm². 3 The cell-rich clusters may comprise at least 5 to 10 times more cells / mg of starting biological material than the dispersed form of the biological material. The method comprises (or substantially comprises, 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 cell-rich clusters from the dispersed form of the biological material; and (e) administering the cell-rich clusters to the mammal. The biological material may be passed through the inlet port and into the conduit without purification of the biological material, without adding any preservatives to the biological material, and without dilution of the biological material. The method may include passing the biological material through the inlet port and into the conduit no more than about 5 minutes after removal of the biological material from the mammal. The method may be one that does not involve centrifugation of the biological material or the cell-rich clusters. Cell-rich clusters can be applied to mammals without exposing the cell-rich clusters to open air. The method may include applying the cell-rich clusters to the mammal within approximately 5 to 30 minutes after obtaining the biological material from the mammal. The method may also include methods that do not involve cultured biological material or cell-rich clusters.

[0011] 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. While similar or equivalent methods and materials to those described herein may be used in the practice of this invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety. In case of conflict, this specification (including definitions) shall prevail. Furthermore, the materials, methods, and embodiments described are illustrative only and not intended to be limiting.

[0012] Details of one or more embodiments of the present invention are set forth in the accompanying drawings and the following description. Other features, objects, and advantages of the invention will become apparent from the specification, the drawings, and the claims. Attached Figure Description

[0013] Figure 1 This is a diagram of an apparatus, according to some embodiments, for collecting biological material from mammals and processing the biological material to form a sample enriched with one or more components of the collected biological material.

[0014] Figure 2 This is a diagram of another apparatus, according to some embodiments, for collecting biological material from mammals and processing the biological material to form a sample enriched with one or more components of the collected biological material.

[0015] Figure 3 This is a diagram of another apparatus, according to some embodiments, for collecting biological material from mammals and processing the biological material to form a sample enriched with one or more components of the collected biological material.

[0016] Figure 4 This is a diagram of another apparatus, according to some embodiments, for collecting biological material from mammals and processing the biological material to form a sample enriched with one or more components of the collected biological material.

[0017] Figure 5 This is a diagram of another apparatus, according to some embodiments, for collecting biological material from mammals and processing the biological material to form a sample enriched with one or more components of the collected biological material.

[0018] Figure 6A and Figure 6B An apparatus according to some embodiments for collecting biological material from mammals and processing the biological material to enrich one or more components of the collected biological material is shown.

[0019] Figure 7 It is a flowchart of a method for collecting biological material from mammals and processing the biological material to enrich one or more components of the collected biological material, according to some implementation schemes.

[0020] Figure 8 It is a flowchart of a method for collecting biological material from mammals and processing the biological material to enrich one or more components of the collected biological material, according to some implementation schemes.

[0021] Figure 9 It is a flowchart of a method for collecting biological material from mammals and processing the biological material to enrich one or more components of the collected biological material, according to some implementation schemes.

[0022] Figure 10 It is a diagram depicting representative components and functions of a peristaltic pump according to some implementation schemes.

[0023] Figure 11 This is a diagram depicting an embodiment of a peristaltic pump having an integrated bottleneck section generated by a spring-loaded element positioned on the outer surface of the conduit.

[0024] Figure 12 This is a diagram depicting an embodiment of a peristaltic pump having an integrated bottleneck section generated by radially movable elements positioned on the pump impeller.

[0025] Figure 13 This is a diagram depicting an embodiment of a peristaltic pump having an integrated bottleneck section formed by a spring-loaded element positioned on the outer surface of the duct and a radially movable roller element positioned on the pump impeller.

[0026] Figure 14 This is a diagram illustrating the apparatus provided herein according to some embodiments, indicating the flow of biological material through the apparatus.

[0027] Figure 15 The figure includes an enlarged view (indicated by the dashed circle and line in the left-hand figure) of a first filtering stage in the apparatus provided herein according to some embodiments.

[0028] Figure 16 The figure includes an enlarged view (indicated by the dashed circle and line in the left-hand figure) of a first filtering stage in the apparatus provided herein according to some embodiments.

[0029] Figure 17 The figure includes an enlarged view (indicated by the dashed circle and line in the upper left figure) of a second filtering stage in the apparatus provided herein according to some embodiments.

[0030] Figure 18 The figure includes an enlarged view (indicated by the dashed circle in the left-hand figure) of the sludge outlet in the apparatus provided herein according to some embodiments.

[0031] In the various figures, the same reference numerals indicate the same elements. Detailed Implementation

[0032] This document provides devices and systems (e.g., bedside devices and systems) designed to receive biological material (e.g., adipose tissue) from a mammal (e.g., a human) and to process the biological material to provide an end product comprising one or more enriched components of the received biological material, which can optionally be reintroduced into the same mammal. For example, this document provides bedside devices designed to collect adipose tissue from a mammal (e.g., a human or a horse) in any suitable setting (e.g., a clinical setting suitable for humans or non-human mammals, such as, but not limited to, a horse) and process the adipose tissue to provide adipose-derived stem cells as an enriched end product of the collected adipose tissue, which can optionally be reintroduced into the same mammal in the same setting (e.g., a clinical setting). This document also provides methods for obtaining biological material (e.g., adipose tissue) from a mammal (e.g., a human) and processing the biological material to provide an end product comprising one or more enriched components of the received biological material, which can optionally be reintroduced into the same mammal. In some cases, the methods presented herein can be used to generate enriched cell populations without centrifugation. Thus, biological material can be obtained from mammals and processed to form, for example, rich clusters of cells, which can be reintroduced into the same mammal without centrifugation.

[0033] In some cases, the final product containing enriched material may contain approximately 1 x 10⁻⁶. 3 To approximately 1 x 10 8 (For example, approximately 7.5 x 10) 4 To approximately 1 x 10 7 Approximately 1 x 10 5 To approximately 2 x 10 5 Approximately 2 x 10 5 To approximately 5 x 10 5 Approximately 5 x 10 5 To approximately 1 x 10 6 Approximately 1 x 10 6 To approximately 5 x 10 6 or approximately 5 x 10 6 To approximately 1 x 10 7 ) cells / cm 3 In some cases, for a specific cell population per milligram of biological material, the enriched product can be enriched 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) compared to the original biological material.

[0034] Any suitable type of cells and other materials may be included in the enriched end product generated using the apparatus and methods provided herein. For example, the enriched end product may contain ADSCs, adipose-derived stromal cells (also known as adipose-derived adult stromal cells or adipose stromal cells), adipocytes, pericytes, preadipocytes, treated liposuction cells, endothelial cells, muscle cells (e.g., smooth muscle cells), fibroblasts, pericytes, vascular pericytes, parietal cells, immune cells (e.g., macrophages, monocytes, lymphocytes, T cells, and / or B cells), hormones, exogenous bodies, purified adipose tissue, erythrocytes, and / or other stem cells (e.g., mesenchymal stem cells). In some cases, the enriched end product generated using the apparatus and methods provided herein may be a stromal vascular portion.

[0035] Generally, the devices provided herein may have a conduit (e.g., a tube) extending from an inlet port to an outlet port. The inlet port may be designed to allow the input of any suitable biological material (e.g., adipose tissue). The inlet port may be designed to have any suitable configuration. For example, the inlet port of the devices provided herein may be designed to connect to the outlet of a liposuction device to collect adipose tissue extracted from a mammal (e.g., a human). In some cases, the inlet port of the devices provided herein may be designed to connect to a Luer connector of a syringe containing biological material (e.g., tissue, such as adipose tissue) obtained from a mammal. The connection between the inlet port of the devices provided herein and another device (e.g., a liposuction device or a syringe) may be any suitable type of connection (e.g., friction fit, press fit, shape fit, and / or material fit connection, such as a clamp, screw, snap, adhesive, or weld connection). The connection may be a detachable connection, a non-detachable connection, and / or a conditionally detachable connection. In some cases, the connection between the inlet port of the device provided herein and another device (e.g., a liposuction device or syringe) capable of providing biological material derived from mammals can be designed to facilitate the aseptic transfer of the biological material into the catheter, ensuring that the biological material is not exposed to contaminants in the external environment. In some cases, the device provided herein may have an adapter located within or adjacent to the inlet port within the catheter, or positioned at the connection between the inlet port of the device and another device (e.g., a liposuction device or syringe) from which the biological material is transferred into the inlet port. In some cases, the adapter may be configured, for example, to facilitate the flow of biological material into the inlet port. For example, if desired, the adapter may be configured to have a shape (e.g., a large funnel-shaped geometry), one or more surface properties (e.g., a microstructured surface, wettability, and / or an applied coating), the ability to adjust viscosity (e.g., by having the ability to add water to reduce viscosity), and / or to introduce internal and / or external forces or pressure differentials to improve the ability to introduce biological material into the device.

[0036] Any suitable biological material can be inserted into the device provided herein (e.g., into a conduit of the device). For example, biological material comprising any suitable 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, vascular pericytes, parietal 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 the device provided herein. In some cases, adipose tissue can be inserted into the device provided herein. In some cases, bone marrow can be inserted into the device provided herein. In some cases, blood can be inserted into the device provided herein.

[0037] In some cases, the biomaterial may be mixed with a fluid (e.g., sterile water or saline) before or during insertion into the inlet port of the device provided herein. Without being limited to any particular mechanism of action, the added fluid may alter (e.g., reduce) the viscosity of the biomaterial, which may facilitate its passage through the conduit of the device provided herein.

[0038] The biomaterials can be derived from any suitable mammal. Examples of mammals from which biomaterials can be obtained for use in the devices provided herein include, but are not limited to, humans, non-human primates (e.g., monkeys), dogs, cats, equines (e.g., horses, donkeys, camels, and zebras), bovids (e.g., cows, bison, and buffalo), pigs, sheep, mice, and rats. In some cases, the biomaterials can be derived from humans (e.g., humans who require stem cell therapy to treat medical conditions such as osteoarthritis, bone / cartilage damage, rheumatic diseases, muscle degeneration, facet joint syndrome, (rotator cuff) tendinopathy, chronic wounds, diabetic foot, alopecia, or multiple sclerosis). In some cases, the biomaterials can be derived from non-human mammals (e.g., horses with damage to their orthopedic musculoskeletal system (such as their articular cartilage and / or tendons), which can be improved through stem cell therapy). Generally, 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 horses, tendon and ligament injuries, osteoarthritis, lameness, degenerative joint changes, wounds, equine metabolic syndrome, digestive diseases, liver diseases, and / or neuromuscular diseases can be treated by removing a biological sample containing stem cells (e.g., adipose tissue), processing the biological sample using the apparatus described herein, and returning the processed biological sample (e.g., a sample enriched with adipose-derived stem cells) to the horse to treat the disease.

[0039] In some cases, the biological material can be obtained directly from mammals, allowing it to be collected directly from the mammal into the inlet port of the device provided herein. For example, when collecting adipose tissue from a mammal (e.g., a human) during a liposuction procedure, the inlet port of the device provided herein can be connected to the outlet of the liposuction device. In some cases, when collecting tissue, bone marrow, or blood samples from a mammal using a syringe connected to a needle (e.g., a biopsy needle or a needle typically used for blood collection), the inlet port of the device provided herein can be directly connected to the Luer connector of the syringe.

[0040] In some cases, biological materials may enter the apparatus provided herein in the same state as when they were extracted from mammals. For example, biological materials may enter the apparatus provided herein without undergoing any purification. For example, biological materials may enter the apparatus provided herein without the addition of any preservatives. For example, biological materials may enter the apparatus provided herein without dilution.

[0041] In some cases, biological materials may enter the device provided herein within approximately 5 minutes of extraction from a mammal. For example, biological materials may enter the device provided herein within approximately 0 to 5 minutes, approximately 0 to 4 minutes, approximately 0 to 3 minutes, approximately 0 to 2 minutes, or approximately 0 to 1 minute after extraction from a mammal. For example, biological materials may enter the device provided herein immediately after extraction from a mammal, such that the biological materials enter the device directly without delay.

[0042] The outlet port may be designed to allow the biological material to exit after it has passed through the catheter and been processed as described herein. The outlet port may be designed to have any suitable configuration. For example, the outlet port of the device provided herein may be designed to connect to a collection vessel. In some cases, the outlet port of the device provided herein may be designed to connect to the inlet of an application device. For example, the outlet port of the device provided herein may be designed to connect to the syringe barrel of a syringe to collect the processed biological material (e.g., a cluster of cells) before transferring it back to the mammal (e.g., from which the original biological material was obtained). In some cases, the outlet port of the device provided herein may be configured to deliver the processed biological material (e.g., a cluster of cells) directly to the mammal (e.g., from which the original biological material was obtained). For example, the outlet port of the device provided herein may be configured such that the obtained processed biological material can be applied directly to the surface to be treated via a spatula and / or nozzle. In some cases, the outlet port of the device provided herein may be configured such that the obtained processed biological material can be directly reinfused into the body via a needle tip.

[0043] In some cases, the outlet port of the device provided herein may be designed to connect to a storage vessel (e.g., a tube or container) in which treated biological material (e.g., a cluster of cells) may be collected and stored. In other cases, the connection between the outlet port of the device provided herein and the collection vessel may be designed to facilitate the aseptic transfer of biological material to the collection container, ensuring that the biological material is not exposed to contaminants in the external environment.

[0044] The conduit of the device provided herein may have any suitable configuration. In some cases, the conduit of the device provided herein may be an elongated tube having an inlet port at one end and an outlet port at the other end. The conduit may have any suitable length. For example, the length of the conduit of the device provided herein from one end to the other may 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).

[0045] The catheters of the devices provided herein can have any suitable cross-sectional shape and size. In some cases, the catheter may have a circular or substantially circular cross-section. In some cases, the catheter may have an elliptical cross-section, a substantially elliptical cross-section, or any cross-sectional region that can be described by a polygon (e.g., a triangle, a rectangle, etc.). In some cases, the cross-sectional shape of the catheter may vary along the length of the catheter. For example, the catheter may 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 elliptical. As described herein, for example, in some cases, the catheter may have a cross-sectional shape along its length that is substantially circular without any applied external force, but is compressed into a more elliptical or elongated shape at the location or in a segment where an external force (e.g., by a dispersing element) is applied.

[0046] The catheter of the device provided herein may have any suitable inner diameter. For example, the inner diameter of the catheter may 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 catheter of the device provided herein may have a maximum inner diameter of not more than about 3 mm, not more than about 5 mm, or not more than about 10 mm at any point along its length. In some cases, the catheter of the device provided herein may have a minimum inner diameter of not less than about 2 mm, not less than about 1.5 mm, not less than about 1 mm, or not less than about 0.5 mm along its entire length, except for one or more segments that may be designed as dispersing elements to disperse biological material into a more dispersed material.

[0047] In some cases, the catheter of the device provided herein may have a constant inner diameter along its entire length. In some cases, the catheter of the device provided herein may include two or more (e.g., two, three, four, five, or more than five) segments, wherein adjacent segments have different inner diameters. For example, the catheter of the device provided herein may have a first segment beginning at an inlet port and a second segment ending at an outlet port, wherein the inner diameter of the second segment is smaller than the inner diameter of the first segment. In some cases, the catheter of the device provided herein may have a first segment with a first inner diameter located downstream of the inlet port and one or more internal segments along the length of the catheter having an inner diameter smaller than the first diameter. In some cases, this smaller inner diameter may be designed to serve as a dispersing element to disperse biological material into a more dispersed material. In some cases, the catheter of the device provided herein may have a first segment beginning at an inlet port, one or more internal segments, and a final segment ending at an outlet port, wherein each segment from the inlet port to the outlet port has a gradually decreasing inner diameter.

[0048] The catheter can be made of any suitable material. In some cases, the catheter may be made of a flexible tube. In some cases, the catheter may include stainless steel, ceramic, glass, thermosetting plastics, thermoplastic polymers, elastomers, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomers, urethanes (e.g., thermoplastic polyurethane), polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber (e.g., natural or synthetic rubber), or any combination thereof.

[0049] The apparatus provided herein may also include one or more pumping elements configured to move biological material through a conduit. For example, the apparatus provided herein may include one, two, three, four, five, or more than five pumping elements. The pumping elements may engage with the conduit to facilitate movement of the biological material through the conduit, such that the biological material moves away from the inlet port and toward the outlet port. Any suitable type of pumping element may be included. For example, the pumping element may be a positive 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, the pumping element may be a direct lifting device (e.g., a reciprocating direct lifting device, or a rotary / continuous direct lifting device). In another example, the pumping element may be a velocity pump (e.g., a reciprocating velocity pump or a rotary / continuous velocity pump), a buoyancy pump, a pulse pump, or a gravity device. In some cases, multiple different types of pumping elements may be included (e.g., two or three different types of pumping elements).

[0050] One or more pumping elements may be located at any suitable position relative to the conduit of the device provided herein. For example, a pumping element may be positioned to contact only the outer portion of the conduit, such that the pumping element is not located within the conduit. In some cases, the entirety of one or more pumping elements may be located outside the conduit, such that no part of any pumping element is located within the conduit. In some cases, the device provided herein may be designed such that no component of any pumping element comes into contact with the biological material within the conduit, whether the biological material is in its original form, its dispersed form, or its treated (e.g., enriched) form.

[0051] In some cases, the apparatus provided herein may include a single pumping element located near the inlet port of the catheter. The pumping element acts to move biological material within the catheter 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, the apparatus provided herein may include more than one pumping element (e.g., two, three, four, five, or more than five pumping elements). In some cases, all pumping elements may be positioned toward the inlet port end of the catheter, upstream of one or more dispersing elements and one or more separating elements. In some cases, two or more pumping elements may be positioned along the length of the catheter (e.g., one or more dispersing elements and one or more separating elements are distributed). In some cases, the apparatus may be configured such that the catheter forms two or more loops (e.g., two, three, four, five, or more than five loops), wherein each loop passes through a pumping element or is otherwise acted upon by a pumping element. For example, each loop may be acted upon by a separate pumping element, or two or more (e.g., all) loops may be acted upon by the same pumping element.

[0052] The apparatus provided herein may also include one or more dispersing elements. For example, the apparatus provided herein may 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 elements may be configured to break up biological material within the conduit, thereby forming a more dispersed form of the biological material. Any suitable type of dispersing element may be included. In some cases, the dispersing element may be an element that utilizes hydrodynamics to disperse the biological material. For example, the conduit of the device provided herein may 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) bottlenecks or constrictions along its length, which reduce the flow opening area of ​​the biomaterial in the conduit. As the biomaterial passes through the bottleneck or constriction, the biomaterial may be broken down, allowing cells within the biomaterial to separate from other material and / or other cells. In some cases, the bottleneck or constriction may be a compression portion of the conduit, wherein this portion is (or has been) compressed by an externally applied force. When a catheter includes two or more compression sections, the compression sections may be compressed in the same direction relative to each other, or in different directions relative to each other. For example, the compression sections may be compressed in the same direction relative to each other such that when the catheter is viewed from the inlet port toward the outlet port (and vice versa), the compression sections appear as flattened regions aligned with each other. Alternatively, the compression sections may be compressed in different directions relative to each other such that when the catheter is viewed from the inlet port toward the outlet port (and vice versa), the compression sections appear as flattened regions at different angles relative to each other. In some cases, the bottleneck or contraction may be a narrowing portion of the catheter material (e.g., a section of tube having a maximum inner diameter smaller than the maximum inner diameter of the adjacent upstream tube section).

[0053] In some cases, the catheter may include two or more bottlenecks or constrictions, wherein each pair of adjacent bottlenecks or constrictions is separated by a mid-section of the catheter. For example, the length of each bottleneck or constriction may be from about 0.05 cm to about 3 cm (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). The mid-section may also have any suitable length. For example, the length of each mid-section may be from about 0.1 cm to about 20 cm (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). In some cases, the devices provided herein may include a catheter configured to form two or more loops (e.g., two, three, four, five, or more than five loops), wherein 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 the catheter forms two or more loops, each loop may pass through or otherwise interact with a pumping element, which facilitates the passage of biological material through the catheter and its bottlenecks or constrictions.

[0054] In some cases, one or more dispersing elements of the devices provided herein may be microstructures formed on the inner surface of the conduit. Examples of microstructures include, but are not limited to, microchannels, plate-like microstructures, and containment elements, including but not limited to a series of baffles made of metal, ceramic, glass, or plastic. The size of the microstructures is typically less than 1 mm.

[0055] In some cases, the dispersing element may not be integral with the catheter. For example, the dispersing element may include a rotor-stator system, an ultrasonic generator, a biochemical solution, or a chemical system. In some cases, the dispersing element may be an active mixer / stirrer (e.g., a stirrer, mixing pump, and / or 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 the dispersing element is an element such as a rotor-stator system or an ultrasonic generator, the dispersing element may be positioned to contact only the outer surface of the catheter, such that the dispersing element is not positioned within the catheter. In some cases, the entirety of one or more dispersing elements may be located outside the catheter, such that no part of any dispersing element is located within the catheter. In some cases, the apparatus provided herein may be designed such that no part of any dispersing element comes into contact with the biological material within the catheter, whether the biological material is in its original form, its dispersed form, or its treated (e.g., enriched) form.

[0056] In some cases, the apparatus provided herein may include a single dispersing element located between at least one pumping element and the outlet port of the catheter. The dispersing element acts to form a more dispersed form of biomaterial within the catheter. In some cases, the apparatus provided herein may include more than one dispersing element (e.g., two, three, four, five, or more than five dispersing elements). In some cases, all dispersing elements may be positioned at least partially centrally along the catheter such that they are downstream of all or more pumping elements and upstream of all or more separating elements. In some cases, two or more dispersing elements may be distributed between two or more pumping elements and one or more separating elements.

[0057] In some cases, the apparatus described herein may have a dispersing element (e.g., a contraction point) directly integrated into a pumping element (e.g., a peristaltic pump) configured to move cellular material through a conduit. Standard peristaltic pumps typically have one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than ten) fixed sliding shoes or rotatable rollers mounted on the pump impeller. One or more sliding shoes or rollers can compress the pump-associated conduit (e.g., a tube) at a pinch point adjacent to the shoe or roller, thereby effectively sealing the conduit. Furthermore, rotation of the pump impeller causes the shoe or roller to move material within the conduit forward through the apparatus.

[0058] In some cases, the apparatus described herein can be configured to combine the delivery function of a pumping element (e.g., a peristaltic pump) with a dispersing element (e.g., a conduit of a compressible peristaltic pump to generate shear at the clamping point, or other element) in a single component. This arrangement can have advantages such as shortening the length of the conduit in the pumping element (which reduces the volume of biomaterial required) and simplifying the machine by integrating pumping and dispersing functions. It should be noted that such an apparatus may have collision problems between the moving roller / slipper and the clamping pin, but these collision problems can be avoided by, for example, using spring-loaded pins, using pins designed to be resilient (e.g., made at least partially of an elastic material (such as rubber, silicone, or thermoplastic elastomers such as thermoplastic polyurethane or thermoplastic polyolefins)), using radially movable or resilient rollers or slippers (e.g., made at least partially of an elastic material (such as rubber or silicone)), or any combination of these methods, against the force applied by the tube. 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 compressing the conduit of the device provided herein (e.g., by pressing against the outer surface of the conduit located distal to the pump impeller) can be periodic and / or reversible, such that each element can be pressed against the conduit for an appropriate period of time (e.g., about 0.5 seconds, about one second, about two to five seconds, or more than five seconds), and then released to allow portions of the conduit previously compressed by the action of the elements to return to an uncompressed configuration. When the 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 apply force against the outer surface of the conduit located proximal to the pump impeller. The compression of the conduit by such radially movable elements can be periodic and / or reversible, such that each element can be pressed against the conduit for an appropriate period of time (e.g., about 0.5 seconds, about one second, about two to five seconds, or more than five seconds) to compress the conduit and assist in dispersing the material contained therein, and also to convey the material through the conduit, and can then be released to allow the portion of the conduit previously compressed by the action of the elements to return to an uncompressed configuration. In some cases, the device provided herein with radially movable elements may also include one or more (e.g., one, two, three, four, five, six, seven, eight, nine, ten, or more than ten) fixing elements that can be pressed against the outer surface of the conduit located distal to the pump impeller, thereby creating a compressed portion (e.g., a bottleneck) in the conduit to assist in dispersing the material contained therein. In some cases, the device provided herein may have a combination of radially movable elements and spring-mounted or elastic elements.

[0059] In some cases, the components that apply force and compress the conduit (e.g., pins and / or other elements) can be continuously pressed against the conduit by forces generated mechanically (e.g., using pre-tensioned springs and / or belts or thin flexible metal sheets with protruding protrusions) or by forces generated by fluids (e.g., pneumatic or hydraulic fluids) and small pneumatic or hydraulic pistons. In some cases (e.g., depending on the material used to manufacture the conduit, the geometry of the conduit, and the desired level of compression), the distance between the pump rollers, slippers, and / or pins may be less than the minimum residual height of the compressed conduit as material passes through. In this case, collisions can be passively avoided by the elastic deformation or radial displacement of the pins and / or rollers and / or slippers. For example, the required deformation energy can be provided by a pump motor. Actively controlled pins and / or rollers and / or slippers can also be used with suitable electromechanical systems. The actuators in such systems can be mechanical, electrical, hydraulic, or pneumatic.

[0060] The apparatus provided herein may include one or more separation elements. For example, the apparatus provided herein may include one, two, three, four, five, or more than five separation elements. One or more separation elements may be configured to separate cells within dispersed biological material from other components of the biological material, thereby forming cell-rich clusters. Any suitable type of separation element may be included. In some cases, the separation elements may form cell-rich clusters based on specific properties of the enriched cells compared to other cells or components within the biological material. For example, the separation elements may separate selected clusters from other cells or components of the biological material based on the size, surface wettability, visual or optical reflectivity, density, inertia, magnetization, or electromobility of the selected cell population.

[0061] In some cases, the separation element may include a filter. Any suitable type of filter can be used. For example, the separation element may include a tangential flow filter (e.g., a hollow fiber filter, a diaphragm filter, or a filter with an integrated filter element manufactured using additive manufacturing). In some cases, the filter may include a filter element of a 3D-printed structure (e.g., a strip) directly integrated into the filter housing, wherein the structure defines a corresponding pore size due to the distance between adjacent structures and the height of the channel. When the separation element includes a filter, the filter may have any suitable pore size. For example, the separation element may include a filter with a pore size of about 1 μm to about 120 μm (e.g., about 1 to about 10 μm, about 10 to about 20 μm, about 20 to about 30 μm, about 30 to about 40 μm, about 40 to about 60 μm, about 60 to about 80 μm, about 80 to about 100 μm, or about 100 to about 120 μm). The pore size can be selected to remove unwanted cellular and / or other components of the biological material from cells and (if any) other materials to be enriched. For example, in some cases, filters with pore sizes of about 50 to about 100 μm (e.g., about 80 μm) can be used to remove cells and other cells (including, for example, adipocytes) larger than about 80 μm from a dispersed form of biological material. In some cases, filters with pore sizes of about 5 to about 15 μm (e.g., about 10 μm) can be used to remove cells and other cells (e.g., fat globules and erythrocytes) smaller than about 10 μm from a dispersed form of biological material. In some cases, the apparatus provided herein may include two or more separation elements, wherein each separation element includes a filter, and wherein the filters have different pore sizes. For example, the apparatus provided herein may include one or more filters as separation elements having a pore size of about 50 to about 100 μm (e.g., about 80 μm) to remove material larger than about 50 to about 100 μm (e.g., about 80 μm) (such as fat cells) from a dispersion of biological material, wherein the resulting material having a size smaller than the pore size advances to one or more additional filters as separation elements having a pore size of about 5 to about 15 μm (e.g., about 10 μm) to remove material smaller than about 5 to about 15 μm (e.g., about 10 μm) (such as fat globules and red blood cells) from a dispersion of biological material, wherein the resulting material having a size larger than the pore size of those additional filters advances toward that outlet port.In some cases, the apparatus provided herein may include one or more filters as separation elements having a pore size of about 5 to about 15 μm (e.g., about 10 μm) to remove material smaller than about 5 to about 15 μm (e.g., about 10 μm) (such as fat globules and erythrocytes) from a dispersion of biological material, wherein the resulting material having a size larger than the pore size advances to one or more additional filters as separation elements having a pore size of about 50 to about 100 μm (e.g., about 80 μm) to remove material larger than about 50 to about 100 μm (e.g., about 80 μm) (such as adipocytes) from a dispersion of biological material, wherein the resulting material having a size smaller than the pore size of those additional filters advances toward that outlet port. One or more separation elements may be located at any suitable position relative to the conduit of the apparatus provided herein. For example, since one or more separation elements are typically in contact with the biological material from which cell populations are to be separated, one or more separation elements may be positioned within the conduit of the apparatus provided herein. When two or more separation elements are included in the apparatus provided herein, the separation elements may be positioned relative to each other in any suitable configuration. For example, two or more separation elements (e.g., two or more filters) may be arranged axially, radially, in parallel, in series, or in combinations of such arrangements (e.g., in series and in parallel) in the device provided herein.

[0062] In some cases, the apparatus provided herein may include a single separating element, wherein the separating element is located downstream of at least one pumping element and at least one dispersing element. In some cases, the apparatus provided herein may include more than one separating element (e.g., two, three, four, five, or more than five separating elements). In some cases, all separating elements may be located downstream of all or all one or more pumping elements and all one or more separating elements. In some cases, two or more separating elements may be distributed with two or more dispersing elements.

[0063] In some cases (e.g., when the device is configured to isolate stem cells), the device provided herein may include at least two stages of filtration. Filters having more than one stage (e.g., two, three, four, or more than four stages) may be generated using any suitable method, including additive manufacturing (also known as 3D printing). Devices including two or more filtration stages may be built into one or more units. For example, the device may include a single separation element (i.e., a single unit) that includes two or more filtration stages. Each of the one or more units may be connected to a conduit such that a flow of material from the conduit may be supplied to and subsequently discharged from the filtration stage. Each filtration stage may 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, the device provided herein may include a first filtration stage with a flow-through filter and a second fine filtration stage with a hollow fiber filter or a tangential flow filter (e.g., with a membrane surface and / or 3D-printed strips) to filter out most particles that are too fine to serve as permeate. Using additive manufacturing, such filtration stages can be produced in a single manufacturing process. In addition, additional functions (such as one or more fastening elements, collection vessels, pressure control elements, vents, and / or elements for controlling material flow) can be integrated simultaneously.

[0064] In some cases, the apparatus provided herein may include one or more (e.g., one, two, three, four, five, or more than five) retention chambers. One or more retention chambers may be in fluid communication with a conduit and may be configured to accumulate and contain biological material (e.g., biological material in its original form, more dispersed form, and / or enriched form). For example, the apparatus provided herein may include one or more retention chambers located along the conduit between an inlet port and a first dispersing element, wherein the one or more retention chambers are configured to accumulate and contain the biological material prior to dispersion. In some cases, the apparatus provided herein may include one or more retention chambers located along the conduit between a first dispersing element and a first separating element, wherein the one or more retention chambers are configured to accumulate and contain a dispersed form of biological material. In some cases, the apparatus provided herein may include one or more retention chambers located along the conduit between a first separating element and an outlet port, wherein the one or more retention chambers are configured to accumulate and contain enriched clusters of cells.

[0065] In some cases, one or more retaining chambers may be connected to the conduit of the device provided herein via press fit, form fit, and / or material fit connections (such as clamps, friction, screws, snaps, adhesives, and / or welded connections). In some cases, the connection may be a detachable connection, a non-detachable connection, or a conditionally detachable connection. In some cases, one or more retaining chambers may be used to smooth the flow of material and / or store material during or after processing (e.g., temporary storage). Material movement may be caused by internal and / or external forces and / or pressure differentials.

[0066] When the apparatus provided herein includes one or more holding chambers, the holding chambers may have any suitable capacity. For example, the holding chamber may be configured to contain about 1 mL to about 3000 mL (e.g., about 1 mL to about 10 mL, about 10 mL to about 50 mL, about 50 mL to about 100 mL, about 100 mL to about 300 mL, about 300 mL to about 500 mL, about 500 mL to about 1000 mL, about 1000 mL to about 2000 mL, or about 2000 mL to about 3000 mL). In some cases, when two or more holding chambers are included, the holding chambers may have the same capacity. In some cases, when two or more holding chambers are included, the holding chambers may have different capacities.

[0067] In some cases, the devices provided herein may include one or more (e.g., one, two, three, four, five, or more than five) inflow ports in fluid communication with a conduit. The one or more inflow ports may be configured to add fluid (e.g., water, saline, or a solution containing one or more cytokines, growth factors, vitamins, hormones, phosphate-buffered saline, culture medium, blood, a solution containing one or more components of adipose tissue or cells, and / or a pharmaceutically active ingredient such as acetylsalicylic acid) to the biomaterial contained within the conduit. When one or more inflow ports are included, they may be located at any suitable location along the conduit between the inlet port and the outlet port. For example, the devices provided herein may include one or more inflow ports fluidly connected to a conduit between the inlet port and a first dispersing element, and / or one or more inflow ports fluidly connected to a conduit between the first dispersing element and a first separating element, and / or one or more inflow ports fluidly connected to a conduit between the first separating element and the outlet port. In some cases, the devices provided herein may lack inflow ports along the conduit between the inlet port and the outlet port.

[0068] Figure 1 To Figure 6 and Figures 10 to 14 E illustrates a representative implementation of the apparatus provided herein.

[0069] refer to Figure 1 The device 100 may be designed to include a conduit 120 having an inlet port 110 and an outlet port 160. The device 100 may also include a pumping element 130, a dispersing element 140, and a separating element 150. The pumping element, dispersing element, and separating element of the device 100 may be located relative to each other at any suitable position. Figure 1 As illustrated in the representative example, the pumping element may be located near the inlet port, the dispersing element may be located downstream of the pumping element, and the separating element may be located downstream of the dispersing element.

[0070] refer to Figure 2 The device 200 may be designed to include more than one pumping element, more than one dispersing element and / or more than one separating element. Figure 2 A representative example of this implementation scheme is shown. For example... Figure 2 As shown, device 200 may 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 may be located relative to each other at any suitable position. Figure 2 As illustrated in the representative example, the pumping element may be located near the inlet port, the dispersing element may be located downstream of the pumping element, and the separating element may be located downstream of the dispersing element.

[0071] refer to Figure 3 The device 300 may be designed to include one or more retaining chambers in fluid communication with the conduit 120. Figure 3 A representative example of this implementation scheme is shown. For example... Figure 3 As shown, the device 300 may have a first retaining chamber 170, a second retaining chamber 172, and a third retaining chamber 174. One or more retaining chambers may be located at any suitable location along the conduit 120. Figure 3 As illustrated in the representative example, a first holding chamber 170 may be located between an inlet port 110 and a pumping element 130, a second holding chamber 172 may be located between a dispersing element 140 and a separating element 150, and a third holding chamber 174 may be located between a separating element 150 and an outlet port 160.

[0072] refer to Figure 4 The device 400 may be designed to include one or more inflow ports that are in fluid communication with the conduit 120. Figure 4 A representative example of this implementation scheme is shown. For example... Figure 4 As shown, the device 400 may have a first inflow port 180 and a second inflow port 185. One or more inflow ports may be located at any suitable location along the conduit 120. Figure 4As illustrated in the representative example, the first inflow port 180 may be located between the pumping element 130 and the dispersing element 140, and the second inflow port 185 may be located between the dispersing element 140 and the separating element 150.

[0073] refer to Figure 5 The device 500 can be designed to include a conduit whose diameter varies along its length. Figure 5 A representative example of this implementation scheme is shown. For example... Figure 5 As shown, the conduit 120 of the device 500 may include a first section 190 and a second section 195, wherein the first section 190 is located upstream of the second section 195, and the diameter of the second section 195 is smaller than the diameter of the first section 190.

[0074] refer to Figure 6A and Figure 6B The device 600 may include a conduit 610 having an inlet port 615 and an outlet port 690, a pumping element 660, and separating elements 680 and 685. The conduit 610 may be flexible and may be configured to form a series of loops 620, 630, 640, and 650. Each loop of the conduit 610 may include a number of compression zones (e.g., compression zones 621, 622, 623, 624, and 625 of loop 620 (where 622-624 are shown as 62x), compression zones 631, 632, 633, 634, and 635 of loop 630, compression zones 641, 642, 643, 644, and 645 of loop 640, and compression zones 651, 652, 653, 654, and 655 of loop 650 (where 652-654 are shown as 65x)). Pumping element 660 may be a peristaltic pump having a series of pump impellers 662, 664, 666, and 668 fixed to shaft 670. Because pump impellers 662, 664, 666, and 668 are fixed to shaft 670, the pump impellers can rotate about shaft 670 at the same speed relative to each other. Loop 620 may be positioned on pump impeller 662, loop 630 may be positioned on pump impeller 664, and loop 640 may be positioned on pump impeller 666, and loop 650 may be positioned on pump impeller 668, such that the rotational action of the pump impellers when shaft 670 rotates can deliver biological material through conduit 610. System 600 may also include separating elements 680 and 685, which may be filters with different pore sizes. For example, separating element 680 may be a filter with a pore size that effectively removes particles larger than the desired cells (e.g., fat particles), while separating element 685 may be a filter with a pore size that effectively removes particles smaller than the desired cells (e.g., blood particles).

[0075] about Figure 10The device 1000 may include a peristaltic pump housing 1040, which may be configured to include a pump impeller 1030, a flexible hollow conduit (e.g., tube 1010) that may contain the material to be conveyed, and one or more fixed slippers or rotatable rollers (e.g., rollers 1020, 1022, 1024, and 1026) mounted on the pump impeller 1030. Rollers 1020, 1022, 1024, and 1026 may effectively compress the tube 1010 at pinch points (e.g., pinch points 1050, 1052, and 1054) adjacent to rollers 1020, 1022, 1024, and 1026, thereby effectively sealing the tube. Furthermore, rotation of the pump impeller 1030 may cause rollers 1020, 1022, 1024, and 1026 to rotate at the pinch points, which may allow material within the tube 1010 to move forward through the tube.

[0076] about Figure 11 The device 1060 can combine the delivery function of a peristaltic pump with a dispersing element including resilient pins and / or spring-mounted pins (e.g., spring-mounted pins 1061, 1062, 1063, 1064, 1065, 1066, and 1067). Regarding... Figure 12 The device 1070 combines the delivery function of a peristaltic pump with radially movable dispersing elements (e.g., radially movable and / or elastic rollers 1071, 1072, 1073, and 1074). The device 1070 may also include stationary dispersing elements (e.g., pins 1081, 1082, 1083, 1084, 1085, 1086, and 1087) that press against the outer surface of the tube 1010 located distal to the pump impeller, thereby creating a compressed portion (e.g., a bottleneck) of the material contained in the tube 1010 for further dispersing. Regarding... Figure 13 The device 1090 can combine the delivery function of a peristaltic pump with dispersing elements, which include combinations of resilient and / or spring-loaded pins (e.g., pins 1061, 1062, 1063, 1064, 1065, 1066, and 1067) and resilient and / or radially movable elements (e.g., rollers 1071, 1072, 1073, and 1074). In some cases, such as Figure 13 As depicted, the apparatus provided herein may 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).

[0077] Figure 14 An implementation scheme for the entire separation element (filter unit) is shown, while Figures 15 to 18 Details of several embodiments of the discrete element are shown.

[0078] about Figure 14 The dashed arrows depict the flow of biological material through a separation element 1100 having a first filtration stage 1130 and a second filtration stage 1160. The separation element 1100 can be connected to a conduit via an inlet connector 1110. Biological material flowing into the first filtration stage 1130 via the inlet connector 1110 can pass through channels in the base plate 1115 and then enter the first filtration stage 1130. Figure 14 As depicted, the flow direction of the elements through the separation unit 1100 is upward or lateral. Regardless of a specific mechanism of action, the upward or lateral movement of the biomaterial facilitates the removal of air bubbles from the material.

[0079] In some cases, the first filtration stage 1130 may 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 discs 1131, 1132, 1133, and 1134). In the first filtration stage 1130, the flow of material may be deflected radially outward from the inlet point from the base plate 1115. The flow may then advance from the outside inward through the stacked filter discs 1131, 1132, 1133, and 1134 into the collection channel 1136, and then upward into the second filtration stage 1160.

[0080] like Figure 14 As depicted, the biological material can flow upward from the first filtration stage 1130 to enter the second filtration stage 1160 through a bottom opening. The material can then flow tangentially across the filter surfaces (e.g., filter membranes 1162 and 1164). The permeate can then be collected from the outside in an annular channel and discharged from the second filtration stage 1160 through outlet 1168.

[0081] The device 1100 may also have a vertical column 1170 fluidly connected to a horizontal conduit 1175, a flushing fluid collection reservoir 1180, and a retentate collection reservoir 1190. Concentrated retentate from the second filtration stage 1160 can be conveyed upwards through the vertical column 1170, which balances the surface tension at small orifices (e.g., approximately 10 µm) in the filtration stage, allowing liquid to pass through the orifices. The retentate can then move into the horizontal conduit 1175, toward the flushing fluid collection reservoir 1180 and the retentate collection reservoir 1190. When the flushing fluid collection reservoir 1180 is full, the material flow of the retentate overcomes the height offset in the horizontal conduit 1175 and reaches the retentate collection reservoir 1190. See also Figure 18 A magnified view is provided for more detail. The prepared biological material (residue) can then be removed from the retention collection container 1190 through the opening 1195. For example, the retention in the retention collection container 1190 can be extracted with a syringe through the opening 1195 and then (directly or later) delivered to the mammal from which the biological sample was initially obtained.

[0082] Figure 15 and Figure 16 Two different design options for the flow-through filter used in the first filtration stage 1130 are depicted. Similarly, dashed arrows depict the flow of biological material through the filter. About Figure 15 Filter 1131 may have a filter housing 1140, which includes a bottom plate 1142, one or more connectors (e.g., connector 1144) for connection to another filter, and a plurality of radial separators (e.g., radial separators 1144). Biomaterial can flow through the radially arranged separators 1144. Figure 15 As depicted, the radial separators can be positioned closer to each other toward the center of the filter 1131, such that the distance between adjacent radial separators decreases toward the center of the filter 1131, and particles that are too large to pass through the filter 1131 are gradually blocked, while smaller particles pass through the first filter unit 1130.

[0083] about Figure 16 Filter 1132 may have a filter housing 1140, which includes a bottom plate 1142, one or more connectors (e.g., connector 1144) for connection to another filter, and a plurality of concentric rings (e.g., rings 1148) having openings for material passage, such that the concentric rings function as separation elements in a flow-through filtration system. Biological material can flow through the concentric rings 1148. Figure 16 As depicted, the openings for biological material to pass through the concentric rings gradually decrease in size from the outside of the filter 1132 toward the center of the filter 1132, so that particles that are too large to pass through the openings in the concentric rings 1148 are gradually blocked, while smaller particles pass through the first filter unit 1130.

[0084] Figure 17 A representative structure of the second filter stage 1160 is illustrated. Regarding... Figure 17 The second filtration stage 1160 may have a central opening 1163, filter membranes 1162 and 1164, a central channel 1165, an outer ring channel 1167, an outlet 1168, and an opening 1169. Biological material can enter the second filtration stage 1160 through the central opening 1163 and then flow tangentially through the filter membranes 1162 and 1164. Due to the hydrostatic pressure of the water column used for retention (e.g., ... Figure 14 (As depicted), most of the liquid material, as permeate, is forced through the pores in filter membranes 1162 and 1164 and guided toward outlet 1168 in outer annular channel 1167. Residue at the end of central channel 1165 passes through opening 1169 in vertical column 1170 (as shown). Figure 14 It is guided upwards in the direction shown.

[0085] Figure 18 The diagram illustrates how a saline or aqueous solution used for rinsing purposes can be separated from the desired retained material via a vertical height offset 1178 in a horizontal conduit 1175. The treated biological material can only enter the retained material collection container 1190 when the rinsing fluid collection container 1180 is filled. The retained material can be temporarily stored in the container 1190 and then removed using, for example, a syringe through opening 1195.

[0086] Generally, the device provided herein can initially be filled with sterile fluid (e.g., saline or sterile water) through the inlet of the conduit, and can pass through the device, through the inlet connector 1110, through the separation element 1100, and upward through the vertical column 1170 to remove air from the device. The fluid can then be removed from the device (e.g., ejected) through biological material placed in the device via the inlet of the conduit. Most sterile fluids (e.g., saline or water) can exit the filter housing via the outlet 1168 (see, e.g., Figure 14 The remaining fluid can then be collected in container 1180. The size of the collection container for flushing fluid and the height of the vertical height offset 1178 ensure that only the desired concentrated material (e.g., stem cells) reaches container 1190.

[0087] This document also provides methods for manufacturing the device provided herein. Any suitable technique can be used to manufacture the device provided herein. In some cases, the device provided herein can be made by positioning one or more separation elements (e.g., filters) into a conduit (e.g., a flexible tube of appropriate length). One or more separation elements can be positioned into the conduit under aseptic conditions, or the device can be sterilized after the separation elements have been positioned into the conduit. In some cases, a first tube section containing an inlet port can be attached to the inflow end of a separation element (e.g., a filter element), and a second tube section can be attached to the outflow end of a separation element (e.g., a filter element). In some cases, the filter element can be designed to allow biomaterial larger than a target size, within a target size (e.g., about 8 to about 80 μm), and smaller than a target size to enter, while excluding (or filtering out) most (e.g., at least 60%, 70%, 80%, 90%, or 95%) of both larger and smaller materials, thereby allowing material within the target size to exit and proceed toward the outlet port. In some cases, additive manufacturing techniques can be used to generate the filter structure (e.g., a porous membrane) used in the device provided herein. For example, the membrane can be 3D printed vertically, or the filaments can be laid horizontally at a desired distance from each other to effectively create pores of a size defined by the distance between adjacent filaments. Methods for manufacturing the device may also include positioning a conduit in contact with one or more pumping elements and one or more dispersing elements. In some cases, one or more inlet ports and / or one or more retaining chambers may be coupled to the conduit such that they are in fluid communication with the conduit. One or more inlet ports and / or retaining chambers may be coupled to the conduit under aseptic conditions, or the device may be sterilized after one or more inlet ports and / or retaining chambers have been coupled to the conduit.

[0088] The apparatus provided herein can be configured to have a first set of articles, such as a power source, a control unit, material handling articles, and fastening articles, and a second set of articles, which may be sterile and disposable. The apparatus can be configured such that biological material comes into direct contact only with the disposable set of articles, thereby enabling feeding, conveying, processing (e.g., dispersion and separation), optional storage, and discharging in a sterile environment. In some cases, the apparatus can ensure that biological material flows from an inlet port through conduits and their components to an outlet port, with the desired biological components being correspondingly concentrated.

[0089] The individual components of a disposable item assembly (e.g., ports, containers, delivery elements, dispersing elements, and separation elements) can be manually assembled to the conduit at the bedside and then secured to the device, or they can be secured to the device as a prefabricated assembly including the conduit. Examples of suitable types of connections for manual assembly include, but are not limited to, press-fit connections, form-fit connections, and / or material-fit connections (such as clamps, friction, screws, snaps, adhesives, and / or welded connections). Such connections can be detachable, non-detachable, or conditionally detachable.

[0090] In some cases, for hygiene reasons, fully assembled prefabricated disposable item sets can be inserted into a machine containing the first set of items, and can be aseptically connected to the same donor / recipient, for example, in a closed system, or to a donor of biological material and a separate recipient.

[0091] This document also provides a method for generating cell-rich clusters from biological material derived from mammals (e.g., humans) using the apparatus provided herein. In some cases, the method provided herein may include inserting biological material from a mammal (e.g., a human) into an inlet port of the apparatus provided herein, such that the biological material can enter a conduit of the apparatus; actuating a pumping element of the apparatus 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 apparatus 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 cell-rich clusters from the dispersed form of the biological material.

[0092] In addition to the steps listed above, the methods provided herein may include one or more optional steps in some cases. For example, the method may include obtaining biological material from a mammal. It should be noted that in some cases, obtaining biological material from a mammal and inserting the biological material into the inlet port of the device provided herein can be completed in a single step, such that the biological material is directly inserted from the mammal into the inlet port. For example, biological material (e.g., adipose tissue) may be obtained using a liposuction device and directly transferred from the liposuction device into the catheter of the device provided herein. In some cases, biological material from a mammal may be obtained in a receiving slot (e.g., a syringe or biopsy needle) and then transferred from the receiving slot into the inlet port and catheter of the device provided herein.

[0093] In some cases, the user can place a second set of sterile disposable items into the device provided herein, and the second set can be secured in the area of ​​the filter housing. The user can then place a catheter into a pumping element (e.g., a peristaltic pump). The entire disposable set can then be flushed (e.g., with saline) to replace the air contained therein with saline. The device can then be connected to a source of biological material (e.g., via...). Figures 1 to 5 The depicted entry point 110) and collection containers can be connected accordingly. In some cases, to ensure a high level of product security in terms of batch tracking and prevention of product piracy, the device provided herein can automatically identify and record inserted disposable sets when the device is equipped with appropriate equipment and has network access; such a device can also query manufacturer approval (e.g., to match barcodes or RFID chips on one or more parts of the disposable set and match the barcodes or RFID chips with the manufacturer's database) and then set machine parameters accordingly. This also ensures that each disposable set is used only once.

[0094] Any suitable type of biological material can be obtained and placed into the device provided herein. Suitable types of biological material include, but are not limited to, adipose tissue, blood, bone marrow, and muscle tissue. Furthermore, any suitable amount of biological material can be placed into the conduit of the device provided herein. For example, about 0.01 L to about 5 L (e.g., about 0.01 L to about 0.05 L, about 0.05 L to about 0.1 L, about 0.1 L to about 0.5 L, about 0.5 L to about 1 L, about 1 L to about 2 L, about 2 L to about 3 L, about 3 L to about 4 L, or about 4 L to about 5 L) of biological material can be inserted into the conduit of the device provided herein and processed to generate cell-rich clusters.

[0095] In some cases, the methods provided herein may include administering an enriched product containing a cluster of cells back to a mammal from which the biological material was obtained. Any suitable amount of the enriched product may be administered. For example, the methods provided herein may include administering about 1 mL to about 500 mL (e.g., about 1 mL to about 10 mL, about 10 mL to about 25 mL, about 25 mL to about 50 mL, about 50 mL to about 100 mL, about 100 mL to about 250 mL, or about 250 mL to about 500 mL) of the enriched product to a mammal. The enriched product may be administered via any suitable route (e.g., by direct injection into a specific organ or tissue, by intravenous injection, by open surgery, by minimally invasive procedures such as arthroscopy or endoscopy, by oral administration, or by topical application).

[0096] In some cases, the method may optionally include adding fluid to the biological material contained within the catheter. For example, the method may include introducing a fluid (e.g., water, saline, or a solution containing one or more cytokines, growth factors, or other additives) into the inlet port of the device provided herein. The fluid may be added at any suitable step of the method provided herein. For example, the fluid may be mixed with the biological material before it is inserted into the inlet port, or the fluid may be introduced into the catheter of the device before or during the dispersion of the biological material contained therein, after dispersion but before the separation of the biological material contained therein, or during the separation of the biological material to generate an enriched product. Any suitable amount of fluid may be introduced and mixed with the biological material. For example, about 0.1 mL to about 2 mL per gram or milliliter of biological material (e.g., about 0.1 mL to about 0.2 mL, about 0.2 mL to about 0.3 mL, about 0.3 mL to about 0.5 mL, about 0.5 mL to about 0.75 mL, about 0.75 mL to about 1 mL, about 1 mL to about 1.5 mL, or about 1.5 mL to about 2 mL) of fluid may be introduced.

[0097] In some cases, the methods described herein may include collecting biological material, a more dispersed form of biological material, and / or rich clusters of cells in one or more retention chambers coupled to a catheter. Movement of the biological material or treated biological material may be caused by internal and / or external forces and / or pressure differences.

[0098] In some cases, the methods provided herein may include generating cell-rich clusters from biological material derived from mammals, and then returning the cell-rich clusters to the mammal. For example, biological material derived from mammals may be transferred from the mammal through the apparatus provided herein and—in its enriched form—returned to the mammal. In some cases, biological material derived from mammals may be transferred from the mammal through the apparatus provided herein and—in its enriched form—directly returned to the mammal, such that the cell-rich clusters are not exposed to open air and / or do not leave the operating room or treatment room where the mammal is located. In some cases, biological material derived from mammals may be transferred from the mammal through the apparatus provided herein and—in its enriched form—directly returned to the mammal, such that the cell-rich clusters are not contaminated.

[0099] In some cases, the methods provided herein may include obtaining biological material from a mammal, processing the biological material using the apparatus provided herein to generate an enriched cell population from the biological material, and reintroducing the enriched cell population into the same mammal within approximately 5 to approximately 30 minutes. For example, biological material may be obtained from a mammal and processed using the apparatus provided herein to generate an enriched cell population, and the enriched cell population may be reintroduced into the same mammal within approximately 5 to approximately 10 minutes, approximately 10 to approximately 15 minutes, approximately 15 to approximately 20 minutes, approximately 20 to approximately 25 minutes, or approximately 25 to approximately 30 minutes.

[0100] In some cases, the methods provided herein may include obtaining biological material from a mammal, processing the biological material using the apparatus provided herein to generate an enriched cell population from the biological material, and reintroducing the enriched cell population into the same mammal, wherein the method does not include a culture step. For example, the biological material may be obtained from a mammal and processed using the apparatus provided herein to generate an enriched cell population, and the enriched cell population may be reintroduced into the same mammal without culturing the biological material or the enriched cell population.

[0101] For example, refer to Figure 7 The method 700 provided herein may include placing biological material from a mammal into a conduit of the device provided herein (710), pumping the biological material through the conduit (720), dispersing the biological material in the conduit (730), separating the dispersed biological material within the conduit to generate a cell-rich cluster (740), and collecting the cell-rich cluster from the conduit (750). In some cases, the method may optionally include collecting the biological material from the mammal (705) and / or administering the collected cells to the mammal (760). It should be noted that the steps within the method may be performed simultaneously. For example, the pumping step 720 may be performed simultaneously with other steps of the method. For example, the pumping step 720 may occur sequentially, such that the biological material is pumped through the conduit while performing the dispersion step 730, the separation step 740, and / or the collection step 750. Additionally, it should be noted that the dispersion step 730, the separation step 740, and the collection step 750 may be performed simultaneously at different points along the conduit, such that different portions of the biological material undergo different steps of the method simultaneously depending on their location within the conduit. Furthermore, it should be noted that the dispersion step 730, the separation step 740, and / or the collection step 750 may be repeated two or more times during the execution of the method. For example, rich clusters of cells may undergo one or more rounds of further dispersion step 730 and / or one or more rounds of further separation step 740 before being collected.

[0102] In some cases, the methods described herein may include one or more steps in which biological material is transferred to a holding chamber. References Figure 8 For example, the method 800 provided herein may include placing biological material from a mammal into a conduit of the 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 the holding chamber (833), returning the dispersed biological material to the conduit (838), separating the dispersed biological material within the conduit to generate a cell-rich cluster (840), and collecting the cell-rich cluster from the conduit (850). In some cases, the method may optionally include collecting the biological material from the mammal (805) and / or administering the collected cells to the mammal (860). Again, it should be noted that the steps within the method may be performed simultaneously. For example, the pumping step 820 may be performed simultaneously with other steps of the method. For example, the pumping step 820 can occur continuously, such that biological material is pumped through the conduit while performing the dispersion step 830, separation step 840, and / or collection step 850. Additionally, it should be noted that the dispersion step 830, separation step 840, and collection step 850 can be performed simultaneously at different points along the conduit, such that different portions of the biological material undergo different steps of the method simultaneously depending on their location within the conduit. Furthermore, it should be noted that the dispersion step 830, separation step 840, and / or collection step 850 can be repeated two or more times during the execution of the method. For example, a rich cluster of cells can undergo one or more rounds of further dispersion step 830 and / or one or more rounds of further separation step 840 before being collected.

[0103] In some cases, the methods provided herein may include one or more steps in which a fluid is added to the biological material. The fluid may be added at any appropriate step, including before or during pumping, before or during dispersion, before or during separation, before or during collection, or any combination thereof. References Figure 9For example, method 900 provided herein may include placing biological material from a mammal into a conduit of the 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 a cell-rich cluster (940), and collecting the cell-rich cluster from the conduit (950). In some cases, the method may optionally include collecting the biological material from the mammal (905) and / or administering the collected cells to the mammal (960). Additionally, it should be noted that steps within the method may be performed simultaneously. For example, the pumping step 920 may be performed concurrently with other steps of the method. For example, the pumping step 920 can occur continuously, such that biological material is pumped through the conduit while performing the dispersion step 930, separation step 940, and / or collection step 950. Additionally, it should be noted that the dispersion step 930, separation step 940, and collection step 950 can be performed simultaneously at different points along the conduit, such that different portions of the biological material undergo different steps of the method simultaneously depending on their location within the conduit. Furthermore, it should be noted that the dispersion step 930, separation step 940, and / or collection step 950 can be repeated two or more times during the execution of the method. For example, a rich cluster of cells can undergo one or more rounds of further dispersion step 930 and / or one or more rounds of further separation step 940 before being collected.

[0104] Exemplary Implementation Implementation 1 is an apparatus for obtaining cell-rich clusters from biological material obtained from mammals, wherein the apparatus comprises: (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to output the cell-rich clusters 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 positioned 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 includes intact cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the cell-rich clusters from the dispersed form of the biological material, wherein the cell-rich clusters per volume comprise a greater number of cells than the dispersed form of the biological material and a less non-cellular material per volume than the dispersed form of the biological material.

[0105] Implementation scheme 2 is the device as described in implementation scheme 1, wherein the rich cluster of cells is a rich cluster of adipose-derived stem cells, mesenchymal stem cells or hematopoietic stem cells.

[0106] Implementation scheme 3 is the device as described in implementation scheme 1 or implementation scheme 2, wherein the cells are adipose-derived stem cells.

[0107] Embodiment 4 is the device as described in any one of Embodiments 1 to 3, wherein the biomaterial comprises adipose tissue.

[0108] Implementation scheme 5 is the apparatus as described in any one of implementation schemes 1 to 4, wherein the mammal is a human.

[0109] Implementation scheme 6 is the device as described in any one of implementation schemes 1 to 4, wherein the mammal is a horse.

[0110] Embodiment 7 is the device as described in any one of Embodiments 1 to 6, wherein the conduit comprises a flexible tube.

[0111] Embodiment 8 is a device as described in any one of Embodiments 1 to 7, wherein the length of the conduit is from about 20 cm to about 10 m.

[0112] Embodiment 9 is a device as described in any one of Embodiments 1 to 8, wherein the conduit includes at least one segment with an inner diameter of about 1 mm to 5 mm.

[0113] Implementation 10 is the device as described in any one of Implementation 1 to 9, wherein the maximum inner diameter at any point along the conduit is not greater than 20 mm.

[0114] Embodiment 11 is the device as described in any one of Embodiments 1 to 10, wherein the minimum inner diameter at any point along the conduit is not less than 0.1 mm.

[0115] Embodiment 12 is the device as described in any one of Embodiments 1 to 11, wherein the conduit comprises stainless steel, ceramic, glass, thermosetting plastic, thermoplastic polymer, elastomer, silicone, polyethylene, polypropylene, polyvinyl chloride, polyurethane, thermoplastic elastomer, urethane polyethersulfone, polysulfone, polyacrylonitrile, polycarbonate, rubber, or any combination thereof.

[0116] Embodiment 13 is the apparatus as described in any one of embodiments 1 to 12, wherein the inlet port is configured to be attached to the outlet of the fat suction device.

[0117] Embodiment 14 is the apparatus as described in Embodiment 13, and further includes an adapter configured to attach the inlet port to the fat suction device.

[0118] Embodiment 15 is an apparatus as described in any one of Embodiments 1 to 12, wherein the inlet port is configured to collect the biological material directly from the mammal.

[0119] Embodiment 16 is the apparatus as described in any one of embodiments 1 to 15, wherein the outlet port is configured to be attached to the inlet of the application device.

[0120] Embodiment 17 is an apparatus as described in any one of embodiments 1 to 15, wherein the outlet port is configured to deliver the rich cluster directly to the mammal.

[0121] Embodiment 18 is an apparatus as described in any one of embodiments 1 to 17, wherein at least one of the one or more pumping elements is a positive displacement pump.

[0122] Implementation scheme 19 is the apparatus as described in implementation scheme 18, wherein the positive displacement pump is a reciprocating pump.

[0123] Implementation scheme 20 is the apparatus as described in implementation scheme 19, wherein the reciprocating pump is a diaphragm pump.

[0124] Embodiment 21 is the apparatus as described in Embodiment 18, wherein the positive displacement pump is a rotary pump or a continuous pump.

[0125] Implementation scheme 22 is the apparatus as described in implementation scheme 21, wherein the positive displacement pump is a peristaltic pump.

[0126] Embodiment 23 is an apparatus as described in any one of embodiments 1 to 22, wherein a component of any one of the one or more pumping elements is not located within the conduit.

[0127] Embodiment 24 is an apparatus as described in any one of embodiments 1 to 23, wherein when the biomaterial and the dispersion of the biomaterial are located within the conduit, components of any one of the one or more pumping elements do not contact the biomaterial or the dispersion of the biomaterial.

[0128] Embodiment 25 is an apparatus as described in any one of embodiments 1 to 24, wherein the one or more dispersing elements are the bottleneck portion of the conduit.

[0129] Implementation scheme 26 is the device as described in implementation scheme 25, wherein the bottleneck is the compression section of the conduit.

[0130] Embodiment 27 is the device as described in Embodiment 25, wherein the conduit includes two or more bottleneck portions separated by a middle section of the conduit, and wherein the bottleneck portions have a reduced maximum diameter compared to the middle section.

[0131] Embodiment 28 is the device as described in any one of embodiments 25 to 27, wherein the conduit includes 5 to 40 bottleneck sections.

[0132] Embodiment 29 is the device as described in any one of embodiments 25 to 27, wherein the conduit includes 20 to 30 bottleneck sections.

[0133] Implementation 30 is the device as described in implementation 29, wherein the conduit is configured as a series of loops.

[0134] Implementation scheme 31 is the apparatus as described in implementation scheme 30, wherein each loop includes 4 to 6 bottleneck sections.

[0135] Implementation 32 is the apparatus as described in Implementation 30 or Implementation 31, wherein each loop is configured to interact with the pumping element.

[0136] Embodiment 33 is an apparatus as described in any one of embodiments 1 to 32, wherein at least one of the one or more dispersing elements comprises one or more microstructures located on the inner surface of the conduit.

[0137] Embodiment 34 is an apparatus as described in any one of embodiments 1 to 33, wherein at least one of the one or more separating elements is a filter.

[0138] Embodiment 35 is the apparatus as described in Embodiment 34, wherein the filter is a tangential flow filter.

[0139] Embodiment 36 is the apparatus as described in Embodiment 35, wherein the tangential flow filter is a hollow fiber filter, a diaphragm filter, or a filter having multiple integrated structures that are separated from each other to define pores.

[0140] Embodiment 37 is an apparatus as described in any one of embodiments 1 to 36, wherein the one or more separating elements are filters arranged in an axial direction.

[0141] Embodiment 38 is an apparatus as described in any one of embodiments 1 to 36, wherein the one or more separating elements are filters arranged radially.

[0142] Embodiment 39 is an apparatus as described in any one of embodiments 1 to 36, wherein the one or more separating elements are filters arranged in parallel.

[0143] Embodiment 40 is an apparatus as described in any one of embodiments 1 to 36, wherein the one or more separating elements are filters arranged in series.

[0144] Embodiment 41 is an apparatus as described in any one of embodiments 1 to 36, wherein the one or more separating elements are filters arranged in series and in parallel.

[0145] Embodiment 42 is an apparatus as described in any one of embodiments 1 to 41, wherein the one or more separation elements include at least one filter with a pore size of about 80 to 100 μm to remove the substance from the enriched cluster in the presence of substances larger than about 80 to 100 μm in the dispersed form of the biomaterial.

[0146] Implementation scheme 43 is the device as described in implementation scheme 42, wherein the larger substance comprises fat cells.

[0147] Embodiment 44 is an apparatus as described in any one of embodiments 1 to 43, wherein the one or more separation elements include at least one filter with a pore size of about 10 μm to remove the substance from the enriched cluster in the presence of substances smaller than about 10 μm in the dispersed form of the biomaterial.

[0148] Implementation scheme 45 is the device as described in implementation scheme 44, wherein the smaller substance includes fat globules, red blood cells, or fat globules and red blood cells.

[0149] Embodiment 46 is an apparatus as described in any one of embodiments 1 to 45, wherein the apparatus includes one or more first retention chambers located along the conduit between the inlet port and a first dispersing element among the one or more dispersing elements, and is configured to accumulate and retain the biomaterial.

[0150] Embodiment 47 is an apparatus as described in any one of embodiments 1 to 46, wherein the apparatus includes one or more second holding chambers located along the conduit between a first dispersing element and a first separating element among the one or more dispersing elements, and is configured to accumulate and retain the dispersed form of the biomaterial.

[0151] Embodiment 48 is an apparatus as described in any one of embodiments 1 to 47, wherein the apparatus includes one or more third retaining chambers located along the conduit between a first separating element and the outlet port of the one or more separating elements, and is configured to accumulate and retain the rich cluster.

[0152] Embodiment 49 is an apparatus as described in any one of embodiments 1 to 48, wherein the apparatus includes one or more inflow ports fluidly connected to the conduit along the conduit located between the inlet port and a first dispersing element of the one or more dispersing elements, and is configured to add fluid to the biomaterial.

[0153] Implementation 50 is the apparatus as described in implementation 49, wherein the fluid is water or salt water.

[0154] Embodiment 51 is an apparatus as described in any one of embodiments 1 to 50, wherein the apparatus includes one or more inflow ports fluidly connected to the conduit along the conduit located between a first dispersing element of the one or more dispersing elements and a first separating element of the one or more separating elements, and is configured to add fluid to the biomaterial.

[0155] Embodiment 52 is the apparatus as described in Embodiment 51, wherein the fluid is water or salt water.

[0156] Embodiment 53 is an apparatus as described in any one of embodiments 1 to 52, wherein the apparatus includes one or more inlet ports fluidly connected to the conduit along the conduit located between a first separation element of the one or more separation elements and the outlet port, and is configured to add fluid to the rich cluster.

[0157] Embodiment 54 is the apparatus as described in Embodiment 53, wherein the fluid is water or salt water.

[0158] Embodiment 55 is an apparatus as described in any one of embodiments 1 to 49, wherein the apparatus lacks any inflow port along the conduit located between the inlet port and the outlet port for fluid connection to the conduit.

[0159] Embodiment 56 is an apparatus as described in any one of embodiments 1 to 55, wherein the apparatus includes no more than one inflow port fluidly connected to the conduit along the conduit located between the inlet port and the outlet port.

[0160] Embodiment 57 is the apparatus as described in any one of embodiments 1 to 56, wherein the rich cluster comprises approximately 150,000 cells / cm². 3 .

[0161] Embodiment 58 is an apparatus as described in any one of embodiments 1 to 57, wherein the rich cluster of cells comprises at least 5 to 10 times more cells / mg of starting biomaterial than the dispersed form of the biomaterial.

[0162] Implementation 59 is an apparatus for obtaining a rich cluster of adipose-derived stem cells from biological material obtained from a mammal and returning the rich cluster to the mammal, wherein the apparatus comprises: (a) a conduit including an inlet port configured to receive the biological material from the mammal and an outlet port configured to deliver the rich cluster 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 positioned 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 includes intact adipose-derived stem cells; and (d) one or more separating elements located downstream of the one or more dispersing elements along the conduit and configured to form the rich cluster of adipose-derived stem cells from the dispersed form of the biological material, wherein the rich cluster of adipose-derived stem cells per volume comprises a greater number of adipose-derived stem cells than the dispersed form of the biological material, and comprises less non-cellular material per volume than the dispersed form of the biological material.

[0163] Implementation 60 is a method for preparing cell-rich clusters using an apparatus as described in any one of Implementations 1 to 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 cell-rich clusters from the dispersed form of the biological material.

[0164] Implementation scheme 61 is the method as described in implementation scheme 60, wherein the biological material is passed through the inlet port and into the conduit without purifying the biological material, adding any preservatives to the biological material, or diluting the biological material.

[0165] Implementation scheme 62 is the method as described in implementation scheme 60 or implementation scheme 61, including allowing the biological material to pass through the inlet port and into the catheter within about 5 minutes after the biological material is removed from the mammal.

[0166] Implementation scheme 63 is a method as described in any one of implementation schemes 60 to 62, wherein the method does not include centrifuging the rich clusters of the biological material or cells.

[0167] Implementation 64 is a method for delivering cells to a mammal using an apparatus as described in any one of Implementations 1 to 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 rich clusters of cells from the dispersed form of the biological material; and (e) applying the rich clusters of cells to the mammal.

[0168] Implementation scheme 65 is the method as described in implementation scheme 64, wherein the biological material is passed through the inlet port and into the conduit without purifying the biological material, adding any preservatives to the biological material, or diluting the biological material.

[0169] Implementation scheme 66 is the method as described in implementation scheme 64 or implementation scheme 65, comprising, no more than about 5 minutes after the biological material is removed from the mammal, passing the biological material through the inlet port and into the catheter.

[0170] Implementation scheme 67 is the method as described in any one of implementation schemes 64 to 66, wherein the method does not include centrifuging the rich clusters of the biological material or cells.

[0171] Implementation scheme 68 is a method as described in any one of implementation schemes 64 to 67, wherein the rich cluster of cells is applied to the mammal without exposing the rich cluster of cells to open air.

[0172] Implementation 69 is a method as described in any one of Implementations 64 to 68, comprising applying the enriched cluster of cells to the mammal within about 5 to about 30 minutes after obtaining the biological material from the mammal.

[0173] Implementation scheme 70 is the method as described in any one of implementation schemes 64 to 69, wherein the method does not include culturing the rich clusters of the biological material or cells.

[0174] The present invention will be further described in the following embodiments, which do not limit the scope of the invention as described in the claims.

[0175] Example Example 1 - Apparatus for collecting human fat and isolating fat-derived stem cells from it The biological material is delivered entirely within a closed system of tubes, filters, reservoirs, and ports, and concentrated according to the desired composition. This closed system is placed in the machine as a single-use item, which can be reused multiple times accordingly.

[0176] The machine is housed in a rollable stainless steel casing, requiring a footprint of approximately 0.4 x 0.4 m and a height of approximately 1.4 m. All surfaces are compatible with standard cleaning and disinfecting agents. Power is limited to normal household power connections. Additionally, WLAN or LAN connectivity is available at the so-called "bedside" location for seamless batch tracking of the sterile, disposable sets used.

[0177] The device is designed to include: • power supply, • Control system, which includes barcode and / or RFID scanners and WLAN / LAN modules. • Drive technology (at least one peristaltic pump having one or more pump impellers, also known as channels), • One or more fastening options, for patient-specific single-use groups. • One or more extrusion points, which are used for flexible tubing in disposable sets, and • One or more additional retainers for use with medical bags and bottles.

[0178] At this device, after capturing the barcode and / or RFID information of the disposable set and releasing the barcode and / or RFID information through a cloud-based database, the disposable set is released for use, and the peristaltic pump is released and preset according to the disposable set.

[0179] The complete assembly (including inlet and outlet ports, a flexible tube as a conduit, at least two separating elements (tangential flow filters, such as hollow fiber filters), and a reservoir) is then attached to the machine. Additionally (e.g., if space permits), the tube is placed in several concentric rings around a single pump impeller (channel) on the peristaltic pump and simultaneously secured in multiple receiving slots. Within these slots, the tube is compressed (e.g., subjected to strong compression) by mechanical tension, causing the circular cross-section to be compressed into a narrow, elongated channel cross-section.

[0180] As the biomaterial is pumped through a flexible tube at a pressure of a few bar by a peristaltic pump, increased shear stress is generated at these extrusion points, which in turn gently disperses the biomaterial. For ease of operation, several clamps are simultaneously locked, each with a movable element. The correct setting of these numerous extrusion points is monitored by sensors in a fault-proofing manner.

[0181] Once the complete assembly is inserted or secured, additional materials are attached to their respective ports. These additional materials can be, for example, additional water or saline solution, but can also be other biological and / or medical materials.

[0182] Once the disposable kit is connected to a material dispenser (e.g., the outlet of a liposuction machine, a bag containing body fat, etc.) and a material receiver (e.g., a receiving bag, syringe, or patient, etc.), the process for enriching the biomaterial is initiated. The biomaterial is continuously and rapidly delivered, dispersed, and concentrated, according to the desired composition (e.g., for the enrichment of adipose-derived mesenchymal stem cells), directly on-site and / or near the patient, in a completely closed, sterile process.

[0183] If the biomaterial is collected directly from mammals (e.g., humans or horses) in the field, the entire process (including the delivery of the enriched biomaterial back to the donor) can be designed to take no more than about 5 to 30 minutes.

[0184] For hygiene reasons, a new, fully prefabricated kit is used for each treatment to reduce any contamination or cross-contamination.

[0185] Example 2 - Apparatus for collecting human fat and isolating fat-derived stem cells from it The biological material is delivered entirely within a closed system of tubes, filters, reservoirs, and ports, and concentrated according to the desired composition. This closed system is placed in the machine as a single-use item, which can be reused multiple times accordingly.

[0186] The machine is housed in a rollable stainless steel casing, requiring a footprint of approximately 0.4 x 0.4 m and a height of approximately 1.4 m. All surfaces are compatible with standard cleaning and disinfecting agents. Power is limited to normal household power connections. Additionally, WLAN or LAN connectivity is available at the so-called "bedside" location for seamless batch tracking of the sterile, disposable sets used.

[0187] The device is designed to include: • power supply, • Control system, which includes barcode and / or RFID scanners and WLAN / LAN modules. • Drive technology (at least one peristaltic pump having one or more pump impellers, also known as channels), • One or more fastening options, for patient-specific single-use groups. • One or more extrusion points, which are used for flexible tubing in disposable sets, and • One or more additional retainers for use with medical bags and bottles.

[0188] At this device, after capturing the barcode and / or RFID information of the disposable set and releasing the barcode and / or RFID information through a cloud-based database, the disposable set is released for use, and the peristaltic pump is released and preset according to the disposable set.

[0189] The complete assembly (including inlet and outlet ports, flexible tubing as conduits, at least two separate stages (integrated in the module or in a separate filter housing), and (if required) a reservoir) is then attached to the machine. The biomaterial flows through at least two filtration stages one after another (e.g., a flow filtration stage and a tangential flow filtration stage). Additionally (e.g., if space permits), the tubing is arranged in several concentric circles around a single pump impeller (channel) on a peristaltic pump and simultaneously secured in multiple receiving slots. Within these slots, the tubing is compressed by mechanical tension, causing the circular cross-section to be compressed into a narrow, elongated channel cross-section.

[0190] As the biomaterial is pumped through the flexible tube at a pressure of a few bar by a peristaltic pump, increased shear stress is generated at these squeeze points, which in turn gently disperses the biomaterial. For ease of operation, several clamps are simultaneously locked, each clamp having a movable element (e.g., a spring-loaded element that contacts the outer surface of the flexible tube located distal to the pump impeller; and / or a radially movable roller element that contacts the outer surface of the flexible tube located proximal to the pump impeller).

[0191] Once the complete assembly is inserted or secured, additional materials are attached to their respective ports. These additional materials can be, for example, additional water or saline solution, but can also be other biological and / or medical materials.

[0192] Once the disposable kit is connected to a material dispenser (e.g., the outlet of a liposuction machine, a bag containing body fat, etc.) and a material receiver (e.g., a receiving bag, syringe, or patient, etc.), the process for enriching the biomaterial is initiated. The biomaterial is continuously and rapidly delivered, dispersed, and concentrated, according to the desired composition (e.g., for the enrichment of adipose-derived mesenchymal stem cells), directly on-site and / or near the patient, in a completely closed, sterile process.

[0193] If the biomaterial is collected directly from mammals (e.g., humans or horses) in the field, the entire process (including the delivery of the enriched biomaterial back to the donor) can be designed to take no more than about 5 to 30 minutes.

[0194] For hygiene reasons, a new, fully prefabricated kit is used for each treatment to reduce any contamination or cross-contamination.

[0195] Example 3 – Treatment of Horses Horses requiring treatment (e.g., horses with tendon injuries) are treated using the device provided herein to promote tendon regeneration. A veterinarian brings the sterile packaged device provided herein, along with a syringe, local anesthetic, and disinfectant, to the location of the horse to be treated (e.g., a veterinary clinic, stable, or even an outdoor paddock or pen). Before beginning to remove the biological material from the animal using the device provided herein (e.g., as described in Embodiment 1 or Embodiment 2), the veterinarian cleans and disinfects the horse's skin. From the time of removal until the concentrated biological material is reintroduced into the horse, the biological material only comes into contact with the original sterile packaged elements.

[0196] Other implementation plans It should be understood that although the invention has been described in conjunction with its detailed description, 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 appended claims.

Claims

1. An apparatus for obtaining cell-rich clusters from biological material derived from mammals, wherein the apparatus 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 rich cluster from the conduit. (b) One or more pumping elements configured to move the biomaterial within the catheter in a direction from the inlet port to the outlet port. (c) One or more dispersing elements, said one or more dispersing elements being positioned along said conduit and configured to form a more dispersed form of said biomaterial from said biomaterial, said dispersed form of said biomaterial comprising intact cells, and (d) One or more separation elements located downstream of the one or more dispersing elements along the conduit and configured to form the cell-rich clusters from the dispersed form of the biomaterial, wherein the cell-rich clusters per volume comprise a greater number of cells than the dispersed form of the biomaterial, and per volume comprise less non-cellular material than the dispersed form of the biomaterial.

2. The device of claim 1, wherein the rich cluster of cells is a rich cluster of adipose-derived stem cells, mesenchymal stem cells or hematopoietic stem cells.

3. The device of claim 1, wherein the cell is an adipose-derived stem cell.

4. The device of claim 1, wherein the biomaterial comprises adipose tissue.

5. The apparatus of claim 1, wherein the mammal is a human.

6. The apparatus of claim 1, wherein the mammal is a horse.

7. The device of claim 1, wherein the conduit comprises a flexible tube.

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

9. The device of claim 1, wherein the conduit includes at least one segment with an inner diameter of about 1 mm to 5 mm.

10. The device of claim 1, wherein the maximum inner diameter at any point along the conduit is not greater than 20 mm.

11. The device of claim 1, wherein the minimum inner diameter at any point along the conduit is not less than 0.1 mm.

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

13. The apparatus of claim 1, wherein the inlet port is configured to be attached to the outlet of the fat suction device.

14. The apparatus of claim 13, further comprising an adapter configured to attach the inlet port to the fat suction device.

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

16. The apparatus of claim 1, wherein the outlet port is configured to be attached to the inlet of the application device.

17. The apparatus of claim 1, wherein the outlet port is configured to deliver the rich cluster directly to the mammal.

18. The apparatus of claim 1, wherein at least one of the one or more pumping elements is a positive displacement pump.

19. The apparatus of claim 18, wherein the positive displacement pump is a reciprocating pump.

20. The apparatus of claim 19, wherein the reciprocating pump is a diaphragm pump.

21. The apparatus of claim 18, wherein the positive displacement pump is a rotary pump or a continuous pump.

22. The apparatus of claim 21, wherein the positive displacement pump is a peristaltic pump.

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

24. The apparatus of claim 1, wherein when the biomaterial and the dispersion of the biomaterial are located within the conduit, a component of any one of the one or more pumping elements does not contact the biomaterial or the dispersion of the biomaterial.

25. The apparatus of claim 1, wherein the one or more dispersing elements are the bottleneck portion of the conduit.

26. The apparatus of claim 25, wherein the bottleneck is a compression section of the conduit.

27. The device of claim 25, wherein the conduit includes two or more bottleneck portions separated by a middle section of the conduit, and wherein the bottleneck portions have a reduced maximum diameter compared to the middle section.

28. The device of claim 25, wherein the conduit comprises 5 to 40 bottleneck sections.

29. The device of claim 25, wherein the conduit comprises 20 to 30 bottleneck sections.

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

31. The apparatus of claim 30, wherein each loop includes 4 to 6 bottleneck sections.

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

33. The apparatus of claim 1, wherein at least one of the one or more dispersing elements comprises one or more microstructures located on the inner surface of the conduit.

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

35. The apparatus of claim 34, wherein the filter is a tangential flow filter.

36. The apparatus of claim 35, wherein the tangential flow filter is a hollow fiber filter, a diaphragm filter, or a filter having a plurality of integrated structures separated from each other to define pores.

37. The apparatus of claim 1, wherein the one or more separating elements are filters arranged axially.

38. The apparatus of claim 1, wherein the one or more separating elements are filters arranged radially.

39. The apparatus of claim 1, wherein the one or more separating elements are filters arranged in parallel.

40. The apparatus of claim 1, wherein the one or more separating elements are filters arranged in series.

41. The apparatus of claim 1, wherein the one or more separating elements are filters arranged in series and in parallel.

42. The apparatus of claim 1, wherein the one or more separating elements comprise at least one filter having a pore size of about 80 to 100 μm to remove the substance from the enriched cluster in the presence of substances larger than about 80 to 100 μm in the dispersed form of the biomaterial.

43. The apparatus of claim 42, wherein the larger substance comprises fat cells.

44. The apparatus of claim 1, wherein the one or more separating elements comprise at least one filter with a pore size of about 10 μm to remove the substance from the enriched cluster in the presence of substances smaller than about 10 μm in the dispersed form of the biomaterial.

45. The device of claim 44, wherein the smaller substance comprises fat globules, red blood cells, or both fat globules and red blood cells.

46. ​​The apparatus of claim 1, wherein the apparatus includes one or more first retaining chambers located along the conduit between the inlet port and a first dispersing element of the one or more dispersing elements, and is configured to accumulate and retain the biomaterial.

47. The apparatus of claim 1, wherein the apparatus includes one or more second holding chambers located along the conduit between a first dispersing element and a first separating element of the one or more dispersing elements, and configured to accumulate and retain the dispersed form of the biomaterial.

48. The apparatus of claim 1, wherein the apparatus includes one or more third retaining chambers located along the conduit between a first separating element and the outlet port of the one or more separating elements, and is configured to accumulate and retain the rich cluster.

49. The apparatus of claim 1, wherein the apparatus includes one or more inflow ports fluidly connected to the conduit along the conduit located between the inlet port and a first dispersing element of the one or more dispersing elements, and is configured to add fluid to the biomaterial.

50. The apparatus of claim 49, wherein the fluid is water or salt water.

51. The apparatus of claim 1, wherein the apparatus includes one or more inflow ports fluidly connected to the conduit along the conduit located between a first dispersing element of the one or more dispersing elements and a first separating element of the one or more separating elements, and is configured to add fluid to the biomaterial.

52. The apparatus of claim 51, wherein the fluid is water or salt water.

53. The apparatus of claim 1, wherein the apparatus includes one or more inflow ports fluidly connected to the conduit along the conduit located between a first separation element of the one or more separation elements and the outlet port, and is configured to add fluid to the rich cluster.

54. The apparatus of claim 53, wherein the fluid is water or salt water.

55. The apparatus of claim 1, wherein the apparatus lacks any inflow port along the conduit located between the inlet port and the outlet port that is fluidly connected to the conduit.

56. The apparatus of claim 1, wherein the apparatus includes no more than one inflow port along the conduit located between the inlet port and the outlet port, which is fluidly connected to the conduit.

57. The apparatus of claim 1, wherein the rich cluster comprises approximately 150,000 cells / cm². 3 .

58. The apparatus of claim 1, wherein the rich cluster of cells comprises at least 5 to 10 times more cells / mg of starting biomaterial than the dispersed form of the biomaterial.

59. An apparatus for obtaining a rich cluster of adipose-derived stem cells from biological material obtained from a mammal and returning the rich cluster to the mammal, wherein the apparatus 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 cluster to the mammal. (b) One or more pumping elements configured to move the biomaterial within the catheter in a direction from the inlet port to the outlet port. (c) One or more dispersive elements, said one or more dispersive elements being positioned along said duct and configured to form a more dispersed form of said biomaterial from said biomaterial, said dispersed form of said biomaterial comprising intact adipose-derived stem cells, and (d) One or more separation elements, located downstream of the one or more dispersion elements along the conduit and configured to form the rich clusters of adipose-derived stem cells from the dispersion of the biological material, wherein the rich clusters of adipose-derived stem cells per volume comprise a greater number of adipose-derived stem cells than the dispersion of the biological material, and comprise less noncellular material per volume than the dispersion of the biological material.

60. A method for preparing cell-rich clusters using the apparatus of claim 1, wherein the method comprises: (a) The biological material of the mammal is passed through the inlet port and into the conduit. (b) Actuating the one or more pumping elements to move the biomaterial within the catheter 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 biomaterial from the biomaterial, and (d) Allowing the one or more separating elements to form the rich clusters of cells from the dispersed form of the biological material.

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

62. The method of claim 60, comprising, no more than about 5 minutes after removing the biological material from the mammal, passing the biological material through the inlet port and into the catheter.

63. The method of claim 60, wherein the method does not include centrifuging the rich clusters of the biological material or cells.

64. A method for delivering cells to a mammal using the apparatus of claim 1, wherein the method comprises: (a) The biological material of the mammal is passed through the inlet port and into the conduit. (b) Actuating the one or more pumping elements to move the biomaterial within the catheter 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 biomaterial from the biomaterial. (d) Allowing the one or more separating elements to form the rich clusters of cells from the dispersed form of the biological material, and (e) The enriched cluster of cells is applied to the mammal.

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

66. The method of claim 64, comprising, no more than about 5 minutes after removing the biological material from the mammal, passing the biological material through the inlet port and into the catheter.

67. The method of claim 64, wherein the method does not include centrifuging the rich clusters of the biological material or cells.

68. The method of claim 64, wherein the rich clusters of cells are applied to the mammal without exposing the rich clusters of cells to open air.

69. The method of claim 64, comprising administering the enriched cluster of cells to the mammal within about 5 to about 30 minutes after obtaining the biological material from the mammal.

70. The method of claim 64, wherein the method does not include culturing the rich clusters of the biological material or cells.

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