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.
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References
Rezanejad, H., Lock, J. H., Sullivan, B. A., & Bonner-Weir, S. (2018). Generation of pancreatic ductal organoids and whole-mount immunostaining of intact organoids. Current Protocols in Cell Biology, 83, e82. https://doi.org/10.1002/cpcb.82.
Zhou, Q., & Melton, D. A. (2018). Pancreas regeneration. Nature., 557, 351–358.
Dor, Y., Brown, J., Martinez, O. I., & Melton, D. A. (2004). Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation. Nature., 429, 41–46.
Xiao, X., Chen, Z., Shiota, C., Prasadan, K., Guo, P., El-Gohary, Y., et al. (2013). No evidence for β cell neogenesis in murine adult pancreas. The Journal of Clinical Investigation, 123, 2207–2217.
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.
Bhartiya, D., Mundekar, A., Mahale, V., & Patel, H. (2014). Very small embryonic-like stem cells are involved in regeneration of mouse pancreas post-pancreatectomy. Stem Cell Research & Therapy, 5, 106.
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.
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.
Ratajczak, M. Z., Ratajczak, J., & Kucia, M. (2019). Very small embryonic-like stem cells (VSELs). Circulation Research, 124, 208–210.
Bhartiya, D., Patel, H., Ganguly, R., Shaikh, A., Shukla, Y., Sharma, D., & Singh, P. (2018). Novel insights into adult and cancer stem cell biology. Stem Cells and Development, 27, 1527–1539.
Lahlil, R., Scrofani, M., Barbet, R., Tancredi, C., & Aries, A. (2018). Hénon. VSELs maintain their pluripotency and competence to differentiate after enhanced ex vivo expansion. Stem Cell Reviews, 14, 510–524.
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.
Monti, M., Imberti, B., Bianchi, N., Pezzotta, A., Morigi, M., Del Fante, C., et al. (2017). A novel method for isolation of pluripotent stem cells from human umbilical cord blood. Stem Cells Development., 26, 1258–1269.
Shaikh, A., Anand, S., Kapoor, S., Ganguly, R., & Bhartiya, D. (2017). Mouse bone marrow VSELs exhibit differentiation into three embryonic germ lineages and germ & hematopoietic cells in culture. Stem Cell Reviews and Reports, 13, 202–216.
Kucia, M., Campbell Reca, R., Zuba-Surma, F. R., Majka, E., Ratajczak, M. J., et al. (2006). A population of very small embryonic-like (VSEL) CXCR4 (+) SSEA-1(+) Oct-4+ stem cells identified in adult bone marrow. Leukemia, 20, 857–869.
White, M. G., Al-Turaifi, H. R., Holliman, G. N., Aldibbiat, A., Mahmoud, A., & Shaw, J. A. (2011). Pluripotency-associated stem cell marker expression in proliferative cell cultures derived from adult human pancreas. The Journal of Endocrinology, 211, 169–176.
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.
Ashizawa, S., Brunicardi, F. C., & Wang, X. P. (2004). PDX-1 and the pancreas. Pancreas., 28, 109–120.
Pedica, F., Beccari, S., Pedron, S., Montagna, L., Piccoli, P., Doglioni, C., & Chilosi, M. (2014). PDX-1 (pancreatic/duodenal homeobox-1 protein 1). Pathologica., 106, 315–321.
Li, D.-S., Yuan, Y.-H., Tu, H.-J., Liang, Q.-L., & Dai, L.-J. (2009). A protocol for islet isolation from mouse pancreas. Nature Protocols, 4, 1649–1652.
Bhartiya, D., Ali Mohammad, S., Guha, A., Pushpa, S., Diksha, S., & Ankita, K. (2019). Evolving definition of adult stem/progenitor cells. Stem Cell Reviews and Reports, 15, 1–3. https://doi.org/10.1007/s12015-019-09879-2.
Zuba-Surma, E. K., Kucia, M., Ratajczak, J., & Ratajczak, M. Z. (2009). “Small stem cells” in adult tissues: very small embryonic-like stem cells stand up! Cytometry. Part A, 75, 4–13.
Chomczynski, P., & Sacchi, N. (2006). The single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroformextraction: twenty-something years on. Nature Protocols, 1(2), 581–585.
Bhartiya, D., Shaikh, A., Anand, S., Patel, H., Kapoor, S., Sriraman, K., Parte, S., & Unni, S. (2016). Endogenous, very small embryonic-like stem cells: critical review, therapeutic potential and a look ahead. Human Reproduction Update, 23(1), 41–76.
Shaikh, A., Bhartiya, D., Kapoor, S., & Nimkar, H. (2016). Delineating the effects of 5-fluorouracil and follicle-stimulating hormone on mouse bone marrow stem/progenitor cells. Stem Cell Research & Therapy, 7, 59.
Hardikar, A. A., Karandikar, M. S., & Bhonde, R. R. (1999). Effect of partial pancreatectomy on diabetic status in BALB/c mice. The Journal of Endocrinology, 162, 189–195.
Anand, S., Bhartiya, D., Sriraman, K., & Mallick, A. (2016). Underlying mechanisms that restore spermatogenesis on transplanting healthy niche cells in busulphan treated mouse testis. Stem Cell Reviews, 12, 682–697.
Yu, K., Fischbach, S., & Xiao, X. (2016). Beta cell regeneration in adult mice: controversy over the involvement of stem cells. 2016. Current Stem Cell Research & Therapy, 11, 542–546.
Domínguez-Bendala, J., Qadir, M. M. F., & Pastori, R. L. (2019). Pancreatic progenitors: there and back again. Trends in Endocrinology and Metabolism, 30(1), 4–11.
Bhartiya, D., & Patel, H. (2015). Very small embryonic-like stem cells are involved in pancreatic regeneration and their dysfunction with age may lead to diabetes and cancer. Stem Cell Research & Therapy, 6, 96–102.
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.
Zanini, C., Bruno, S., Mandili, G., Baci, D., Cerutti, F., Cenacchi, G., Izzi, L., Camussi, G., & Forni, M. (2011). Differentiation of mesenchymal stem cells derived from pancreatic islets and bone marrow into islet-like cell phenotype. PLoS One, 6, e28175.
Bhartiya, D. (2013). Are mesenchymal cells indeed pluripotent stem cells or just stromal cells? OCT-4 and VSELs biology has led to better understanding. Stem Cells International, 2013, 547501.
Clevers, H., & Watt, F. M. (2018). Defining adult stem cells by function, not by phenotype. Annual Review of Biochemistry, 87, 1015–1027.
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.
Cho, J., D'Antuono, M., Glicksman, M., Wang, J., & Jonklaas, J. (2018). A review of clinical trials: mesenchymal stem cell transplant therapy in type 1 and type 2 diabetes mellitus. American Journal of Stem Cells, 7(4), 82–93.
Sipp, D., Robey, P. G., & Turner, L. (2018). Clear up this stem-cell mess. Nature, 561, 455–457.
Caplan, A. I. (2019). There is no “stem cell mess”. Tissue Engineering. Part B, Reviews, 25, 291–293. https://doi.org/10.1089/ten.TEB.2019.0049.
Banakh, I., Gonez, L. J., Sutherland, R. M., Naselli, G., & Harrison, L. C. (2012). Adult pancreas side population cells expand after β cell injury and are a source of insulin-secreting cells. PLoS One, 7, e48977.
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.
Berrocal, T., Luque, A. A., Pinilla, I., & Lassaletta, L. (2005). Pancreatic regeneration after near-total pancreatectomy in children with nesidioblastosis. Pediatric Radiology, 35(11), 1066–1070.
Menge, B. A., Tannapfel, A., Belyaev, O., Drescher, R., Müller, C., Uhl, W., et al. (2008). Partial pancreatectomy in adult humans does not provoke β-cell regeneration. Diabetes, 57, 142–149.
Menge, B. A., Breuer, T. G., Ritter, P. R., Uhl, W., Schmidt, W. E., & Meier, J. J. (2012). Long-term recovery of β-cell function after partial pancreatectomy in humans. Metabolism., 61, 620–624.
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.
Swartz, F. J., & Carstens, P. H. (1986). An islet of Langerhans located within the epithelium of a human pancreatic duct. Histology and Histopathology, 1, 111–117.
Tainaka, K., Kubota, S. I., Suyama, T. Q., Susaki, E. A., Perrin, D., Ukai-Tadenuma, M., Ukai, H., & Ueda, H. R. (2014). Whole-body imaging with single-cell resolution by tissue decolorization. Cell., 159, 911–924.
Bhartiya, D. (2019). Clinical translation of stem cells for regenerative medicine a comprehensive analysis. Circulation Research, 124, 840–842.
Shapiro, A. M. J. (2018). Gearing up for stem cell-derived beta cells-are we ready? Transplantation, 102(8), 1207–1208.
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.
Pepper, A. R., Bruni, A., Pawlick, R., O'Gorman, D., Kin, T., Thiesen, A., & Shapiro, A. M. J. (2019). Posttransplant characterization of long-term functional hESC-derived pancreatic endoderm grafts. Diabetes., 68(5), 953–962.
Pawitan, J. A., Yang, Z., Wu, Y. N., & Lee, E. H. (2018). Towards standardized stem cell therapy in type 2 diabetes mellitus: a systematic review. Current Stem Cell Research & Therapy, 13(6), 476–488.
Bhartiya, D. (2017). Pluripotent stem cells in adult tissues: struggling to be acknowledged over two decades. Stem Cell Reviews, 13(6), 713–724.
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.
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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.
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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.
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Part of the work was presented as a Poster at ISSRF Annual Meeting held in New Delhi in February 2019.
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Mohammad, S.A., Metkari, S. & Bhartiya, D. Mouse Pancreas Stem/Progenitor Cells Get Augmented by Streptozotocin and Regenerate Diabetic Pancreas After Partial Pancreatectomy. Stem Cell Rev and Rep 16, 144–158 (2020). https://doi.org/10.1007/s12015-019-09919-x
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DOI: https://doi.org/10.1007/s12015-019-09919-x