Evaluating KIND1 human embryonic stem cell-derived pancreatic progenitors to ameliorate streptozotocin-induced diabetes in mice

Abstract

Background & objectives: Diabetes is a global disease burden. Various stem cell types are being explored to serve as an alternative source of islets. This study was conducted to evaluate the ability of in-house developed human embryonic stem (hES) cells-derived pancreatic progenitors to ameliorate diabetic symptoms in mice.

Methods: Pancreatic progenitors were packed in macro-capsules and transplanted into six male Swiss mice and four mice were taken as controls. Thirty days post-transplantation, diabetes was induced by streptozotocin treatment. Mice were then followed up for >100 days and body weight and blood glucose levels were regularly monitored.

Results: Control mice lost weight, maintained high glucose levels and did not survive beyond 40 days, whereas transplanted group maintained body weight and four of the six mice had lowered blood glucose levels. About five-fold increase was observed in human C-peptide levels in the recipients of progenitor transplants as compared to diabetic control.

Interpretation & conclusions: The beneficial effect of transplanted cells was not long-lasting. Further studies are required to critically evaluate and compare the potential of endogenous pluripotent stem cells and hES cells-derived progenitors before moving from bench to the bedside.

Fig. 1

Human embryonic stem cells (KIND1) used for the study. (A & B) KIND1 human embryonic stem cells colony grown as monolayer under feeder-free conditions on a Geltrex-coated dish (×10). (C) Pluripotent transcripts amplified by reverse transcription polymerase chain reaction using KIND1 human embryonic stem cells OCT4 (octamer-binding transcription factor 4), NANOG, SOX2 (SRY-Box 2), REX1 (Reduced Expression Protein 1) and TERT (Telomerase Reverse Transcriptase). GAPDH was used as housekeeping gene. (D) Immunofluorescence staining revealed the localization of markers such as OCT4, NANOG and SSEA4 (stage-specific embryonic antigen-4) (×40) (top & middle panel) and ×20 (bottom panel). Counterstaining was done using PI for OCT4 while DAPI was used for NANOG and SSEA4.

Fig. 2

Differentiation of KIND1 cells into pancreatic progenitors. (A) Schematic representation of protocol. (B) Expression of pluripotency (OCT4, SOX2), pancreatic progenitor-specific (SOX17 (SRY-Box 17), SOX9 (SRY-Box 9), NKX6.1 (NK6 homeobox 1), PDX1 (Pancreas/duodenum homeobox protein 1), ectodermal SOX1 (SRY-Box 1), MAP2 (Microtubule-associated protein 2) and mesodermal MESP1 (mesoderm posterior bHLH transcription factor 1), NKX2.5 (NK2 homeobox 5) transcripts in undifferentiated (UD, blue) and pancreatic progenitors on day 16 (PP, red). Data represent mean±standard error of the mean. (C) Immunofluorescence for SOX9, SOX17 and PDX1 (×10).