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Mouse Pancreas Stem/Progenitor Cells Get Augmented by Streptozotocin and Regenerate Diabetic Pancreas After Partial Pancreatectomy

  • Subhan Ali Mohammad
  • Siddhanath Metkari
  • Deepa BhartiyaEmail author
Article
  • 40 Downloads

Abstract

Existence of stem cells in adult pancreas remains contentious. Single cells suspensions obtained by collagenase and trypsin digestion separately from adult mouse pancreas and pancreatic islets were spun at 1000 rpm (250 g) to collect the cells. At this speed the stem/ progenitor cells remained buoyant and were further enriched by spinning the supernatant at 3000 rpm (1000 g). Two distinct populations of stem cells were detected including pluripotent, very small (2–6 μm) embryonic-like stem cells (VSELs) that expressed nuclear OCT-4A and pluripotent transcripts (Oct-4A, Sox2, Nanog, Stella) and slightly bigger progenitors, pancreatic stem cells (PSCs) that expressed cytoplasmic OCT-4B and PDX-1. Streptozotocin treated diabetic pancreas showed an increase in numbers of VSELs (2–6 μm, 7AAD-, LIN-CD45-SCA1+ cells) and up-regulation of transcripts specific for stem/ progenitor cells. Diabetic mice were further subjected to partial pancreatectomy to study involvement of VSELs/ PSCs during regeneration. VSELs/ PSCs were mobilized in large numbers, were observed in the lumen of blood vessels and PCNA expression suggested their proliferation. Initially, new acini assembled to regenerate the exocrine pancreas and later by Day 30, neogenesis of islets was observed in the vicinity of the blood vessels and pancreatic ducts by the differentiation of endogenous VSELs/ PSCs which may be targeted to regenerate diabetic pancreas in clinical settings.

Keywords

Pancreas Stem cells VSELs Diabetes Regeneration PDX-1 OCT-4 

Notes

Acknowledgements

Thanks to Ms. Pushpa Singh and Ms. Ankita Kaushik for their help. Authors acknowledge the help provided by Drs Ramesh Bhonde (Pune) and Shahir Gaikwad (Department of Surgery, Bombay Veterinary College, Mumbai) towards surgical manipulations of mouse pancreas. Sincere thanks to Dr. Aleem Khan (Deccan College of Medical Sciences, Hyderabad) for his help to extract good quality RNA from pancreas for qRT-PCR studies. Thanks to the central facilities at NIRRH of Confocal Microscopy and Flow Cytometry for their expert help. Authors also thank Shri Vaibhav Shinde for help with the art work. DB is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Author Contributions

SAM has done all the work, data interpretations and helped with manuscript preparation. SMM performed all the surgeries involved in the study and participated in the discussions. DB conceptualized the project, arranged the funds, helped with study design data, interpretation and manuscript preparation. All the authors critically reviewed the manuscript and provided intellectual inputs.

Funding Information

Thanks to ICMR Centenary PDF Scheme (3/1/3/PDF [16]/−2017-HRD-5) to support this work at NIRRH, Mumbai. NIRRH Accession Number RA/755/03–2019.

Compliance with Ethical Standards

Conflict of Interest

No potential conflicts of interest relevant to this article were reported.

Supplementary material

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ESM 1 (PDF 1310 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Stem Cell Biology DepartmentICMR- National Institute for Research in Reproductive HealthMumbaiIndia
  2. 2.Experimental Animal FacilityICMR-National Institute for Research in Reproductive HealthMumbaiIndia

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