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

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. 20 24 20 46 58 04 J ul 2 02 4 1 0 0 5 3 7 8 1 6 7 A B S T R A C T 2 0 2 4 2 0 4 6 5 8 0 4 J u l 2 0 2 4

Claims

1. A method of generating an activated islet proliferating cell (AIPC) population, comprising:culturing a population of pancreatic islets in vitro with a cell culture medium comprising a base medium and an effective amount of an active agent to obtain the AIPC population,wherein at least 60% of the AIPC population is comprised of a CD133 cell marker and a Ki-67 cell marker;wherein at least 60% of the AIPC population is capable of insulin production; and wherein the active agent comprises a polypeptide having at least 50% sequence identity to any one of SEQ ID NOs: 1 -4.

2. The method of claim 1, wherein the population of pancreatic islets comprise from about 10 to about 10,000 islets.

3. The method of any one of the preceding claims, wherein the culturing occurs for a period of about 2 to about 10 days.

4. The method of any one of the preceding claims, wherein at least about 80% of the AIPC population is positive for insulin, CD133, and Ki-67.

5. The method of any one of the preceding claims, wherein the polypeptide has at least: 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1,99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, the amino acid sequence set forth in SEQ ID NO: 1, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, the amino acid sequence set forth in SEQ ID NO: 2, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 3, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 3, 99% sequence identity to the amino acid sequence set 342024204658   04 Jul 2024forth in SEQ ID NO: 3, the amino acid sequence set forth in SEQ ID NO: 3, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4.

6. The method of any one of the preceding claims, wherein the cell culture medium comprises the active agent at a concentration of about 1 pg / mL to about 20 pg / mL.

7. The method of any one of the preceding claims, wherein the cell culture medium comprises the active agent at a concentration of about 5 pg / mL to about 15 pg / mL.

8. A composition comprising an activated islet proliferating cell (AIPC) population, wherein 60% of the AIPC population is comprised of a CD133 cell marker and a Ki-67 cell marker, andwherein at least 60% of the AIPC population is capable of insulin production.

9. The composition of claim 8, wherein the AIPC population is capable of serial passage and remain viable for over 60 days in culture.

10. The composition of any one of claims 8-9, further comprising one or more of a buffer, one or more of a pharmaceutically acceptable carrier, or one or more of a pharmaceutically acceptable additive.

11. A composition comprising an activated islet proliferating cell (AIPC) population obtained by a process comprising:culturing an isolated pancreatic cell population in vitro with a cell culture medium comprising a base medium and an effective amount of an active agent to obtain the AIPC population,wherein at least 30% of the AIPC population is comprised of a CD133 cell marker and a Ki-67 cell marker;2024204658   04 Jul 2024wherein at least 60% of the AIPC population is capable of insulin production; and wherein the active agent comprises a polypeptide having at least 50% sequence identity to any one of SEQ ID NOs: 1 -4.

12. The composition of claim 12, wherein the AIPC population is capable of serial passage and remains viable for at least 60 days in culture.

13. The composition of any one of claims 12-13, wherein the population of pancreatic islets comprise from about 10 to about 10,000 islets.

14. The composition of any one of claims 12-14, wherein the culturing occurs for a period of about 2 to about 10 days.

15. The composition of any one of claims 12-15, wherein at least about 80% of the AIPC population is positive for insulin, CD133, and Ki-67.

16. The composition of any one of claims 12-16, wherein the polypeptide has at least: 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 1, the amino acid sequence set forth in SEQ ID NO: 1, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, 98% sequence identity to the amino acidsequence set forth in SEQ ID NO: 2, 99% sequence identity to the amino acidsequence set forth in SEQ ID NO: 2, the amino acid sequence set forth in SEQ ID NO:2, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 3, 98%sequence identity to the amino acid sequence set forth in SEQ ID NO: 3, 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 3, the amino acid sequence set forth in SEQ ID NO: 3, 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, 98% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, 99% sequence identity to the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID2024204658   04 Jul 2024NO: 4.

17. The composition of any one of claims 12-17, wherein the cell culture medium comprises the active agent at a concentration of about 1 pg / mL to about 20 pg / mL or about 5 pg / mL to about 15 pg / mL.

18. A composition according to any one of claims 8-17 for use in the treatment of type I diabetes.

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