Mouse Pancreas Stem/Progenitor Cells Get Augmented by Streptozotocin and Regenerate Diabetic Pancreas After Partial Pancreatectomy
- 2 Downloads
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.
KeywordsPancreas Stem cells VSELs Diabetes Regeneration PDX-1 OCT-4
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.
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.
Thanks to ICMR Centenary PDF Scheme (3/1/3/PDF /−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.
- 5.Qadir, M. M. F., Álvarez-Cubela, S., Klein, D., Lanzoni, G., García-Santana, C., Montalvo, A., Pláceres-Uray, F., Mazza, E. M. C., Ricordi, C., Inverardi, L. A., Pastori, R. L., & Domínguez-Bendala, J. (2018). P2RY1/ALK3-expressing cells within the adult human exocrine pancreas are BMP-7 expandable and exhibit progenitor-like characteristics. Cell Reports, 22, 2408–2420.PubMedPubMedCentralCrossRefGoogle Scholar
- 7.Zuba-Surma, E. K., Kucia, M., Wu, W., Klich, I., Lillard, J. W., Jr., Ratajczak, J., & Ratajczak, M. Z. (2008). Very small embryonic like stem cells are present in adult murine organs: image stream based morphological analysis and distribution studies. Cytometry., 73, 1116–1127.CrossRefGoogle Scholar
- 8.Starzyńska, T., Dąbkowski, K., Błogowski, W., Zuba-Surma, E., Budkowska, M., Sałata, D., Dołęgowska, B., Marlicz, W., Lubikowski, J., & Ratajczak, M. Z. (2013). An intensified systemic trafficking of bone marrow-derived stem/progenitor cells in patients with pancreatic cancer. Journal of Cellular and Molecular Medicine, 17, 792–799.PubMedPubMedCentralCrossRefGoogle Scholar
- 12.Havens, A. M., Sun, H., Shiozawa, Y., Jung, Y., Wang, J., Mishra, A., Jiang, Y., O'Neill, D. W., Krebsbach, P. H., Rodgerson, D. O., & Taichman, R. S. (2014). Human and murine very small embryonic-like cells represent multipotent tissue progenitors, in vitro and in vivo. Stem Cells and Development, 23, 689–701.PubMedCrossRefGoogle Scholar
- 17.Zhao, M., Amiel, S. A., Christie, M. R., Muiesan, P., Srinivasan, P., Littlejohn, W., Rela, M., Arno, M., Heaton, N., & Huang, G. C. (2007). Evidence for the presence of stem cell-like progenitor cells in human adult pancreas. The Journal of Endocrinology, 195, 407–414. https://doi.org/10.1677/JOE-07-0436.CrossRefPubMedGoogle Scholar
- 31.Karaoz, E., Ayhan, S., Gacar, G., Aksoy, A., Duruksu, G., Okçu, A., Demircan, P. Ç., Sariboyaci, A. E., Kaymaz, F., & Kasap, M. (2010). Isolation and characterization of stem cells from pancreatic islet: pluripotency, differentiation potential and ultrastructural characteristics. Cytotherapy., 12, 288–302.PubMedCrossRefGoogle Scholar
- 35.Furuyama, K., Chera, S., van Gurp, L., Oropeza, D., Ghila, L., Damond, N., Vethe, H., Paulo, J. A., Joosten, A. M., Berney, T., Bosco, D., Dorrell, C., Grompe, M., Ræder, H., Roep, B. O., Thorel, F., & Herrera, P. L. (2019). Diabetes relief in mice by glucose-sensing insulin-secreting human α-cells. Nature., 567(7746), 43–48.PubMedPubMedCentralCrossRefGoogle Scholar
- 40.Augstein, P., Loudovaris, T., Bandala-Sanchez, E., Heinke, P., Naselli, G., Lee, L., Hawthorne, W. J., Góñez, L. J., Neale, A. M., Vaillant, F., Thomas, H. E., Kay, T. W., Banakh, I., & Harrison, L. C. (2018). Characterization of the human pancreas side population as a potential reservoir of adult stem cells. Pancreas., 47, 25–34.PubMedCrossRefGoogle Scholar
- 44.Wang, P., Karakose, E., Liu, H., Swartz, E., Ackei, C., Zlatanic, V., et al. (2018). Combined inhibition of DYRK1A, SMAD, and Trithorax pathways synergizes to induce robust replication in adult human beta cells. Cell Metabolism S1550–4131; 18:30742–3.Google Scholar
- 49.Odorico, J., Markmann, J., Melton, D., Greenstein, J., Hwa, A., Nostro, C., Rezania, A., Oberholzer, J., Pipeleers, D., Yang, L., Cowan, C., Huangfu, D., Egli, D., Ben-David, U., Vallier, L., Grey, S. T., Tang, Q., Roep, B., Ricordi, C., Naji, A., Orlando, G., Anderson, D. G., Poznansky, M., Ludwig, B., Tomei, A., Greiner, D. L., Graham, M., Carpenter, M., Migliaccio, G., D’Amour, K., Hering, B., Piemonti, L., Berney, T., Rickels, M., Kay, T., & Adams, A. (2018). Report of the key opinion leaders meeting on stem cell-derived beta cells. Transplantation., 102(8), 1223–1229.PubMedPubMedCentralCrossRefGoogle Scholar