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Compositions and methods for propagating insulin-producing islet cells and therapeutic uses thereof

A composition, insulin technology, used in biochemical devices and methods, pancreatic cells, cell culture supports/coatings, etc.

Pending Publication Date: 2021-06-22
想象制药有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While this work appears promising, the level of proliferation observed is insufficient to generate the number of insulin-producing cells necessary for a living cell source of insulin-producing cells for therapeutic use, such as for the treatment of diabetes

Method used

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  • Compositions and methods for propagating insulin-producing islet cells and therapeutic uses thereof
  • Compositions and methods for propagating insulin-producing islet cells and therapeutic uses thereof
  • Compositions and methods for propagating insulin-producing islet cells and therapeutic uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0075] In an exemplary study, 300 murine islets were isolated from pancreases of 8- to 10-week-old male BALB / c mice (eg, from Jackson Labs or Charles River). In CMRL-containing medium (originally developed by ConnaughtMedical Research Laboratories and currently available from Mediatech et al.) Isolated islets were cultured in culture medium. Treated cells were grown and compared with control cells, which were the same cells grown in basal medium without IMG. Within 24 hours, fewer cells were observed growing in cell culture medium containing IMG based on visual inspection at 10x magnification through a microscope (using a Leica light microscope with Lumenera camera, Infinity analysis software) clusters, and cultured islet cells appear to actively migrate away from cell clusters. On the other hand, control islet cells were observed to maintain their cluster morphology and show minimal signs of cell migration. Three days after treatment, islet cells cultured in IMG-containing...

Embodiment 2

[0078] In another exemplary study, human islets were used. Human islets, such as Human Islets for Research It is primary human islets processed from organ donor pancreases, which are commercially available from Prodo Laboratories, Incorporated. Typically, the human islet population is less than 3% positive for Ki-67, and healthy islets are about 50% insulin-producing in response to stimulation. In this example, ten (10) islets were cultured in CMRL 1066 medium, 10% fetal bovine serum, 10% human serum, 2 mmol / L-glutamine, antibiotics, and IMG at a concentration of about 10.0 μg / mL. cultured in culture medium. Islets cultured in medium containing IMG were comparable to control cells cultured in CMRL 1066 medium without IMG (alone). Treated and untreated human islets were cultured at 37°C for 5 days prior to CyQUANT cell proliferation assay (eg, from Invitrogen). Within 5 days, human islets cultured in medium containing IMG showed approximately 1.5 times the number of cells ...

Embodiment 3

[0080] Separately, human islets (3,000-10,000) were cultured in control medium or medium containing CMRL 1066 supplemented with 10% fetal bovine serum, 10% human serum, 2mmol / L-glutamine, antibiotics and 10.0μg / ml concentration In the culture medium of IMG, the anchorage factor mixture (AFM ) plate culture. Different ratios of ECAF and collagen can be used, including a 50 / 50 ratio of collagen and ECAF. A thin layer of AFM (between 3-10 mL) was applied and after 30 minutes of settling, excess AFM was removed and the plate was allowed to dry in the laminar bench for 45 minutes. Before use, wash the plate with PBS to remove any potential contamination. Cultured islets are grown in AFM-treated flasks (plates can be used) in medium or control medium for 2 to 5 days.

[0081] Based on observations using bright-field microscopy, control islets (cultured in standard CMRL medium without IMG) appeared to maintain their cluster morphology and showed no signs of cell migration. The ex...

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Abstract

Disclosed herein are compositions and methods to differentiate pancreatic cells into functional insulin-producing CD133 / Ki-67-positive activated islet proliferating cells (AIPCs) derived from isolated pancreatic islets and expand derived-AIPCs in in vitro cultures using a culture medium comprising an active agent. Also disclosed herein is the use of the AIPCs for implantation into a mammal for in vivo therapy, specifically for pancreatic disorders, including diabetes type I.

Description

[0001] References to related applications [0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 689,780, filed June 25, 2018, which is incorporated herein by reference in its entirety. Background technique [0003] Diabetes is a metabolic disease defined by high blood sugar, but it also causes serious damage to a variety of body systems, including nerves, blood vessels, eyes, kidneys, heart, and more. Some studies estimate that there may be more than 400 million people alive with diabetes worldwide, and the incidence and incidence of diabetes is increasing globally, especially in the United States, China and India. [0004] There are many variants of diabetes, but two main types predominate. In type 1 diabetes, the pancreas produces little or no insulin due to the loss of insulin-producing pancreatic beta-cells. These insulin-producing islet cells are present in type 2 diabetes, but the amount and effectiveness of insulin produced is lowe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/03A61K35/39C12N5/071A61P5/50A61P3/10
CPCA61P5/00A61K35/39C12N5/0678C12N2501/998C12N5/0677C12N2533/54A61P5/48
Inventor 恩戈克·泰乔纳森·波莱特
Owner 想象制药有限责任公司
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