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Extrinsic Factors Promoting In Vitro Differentiation of Insulin-Secreting Cells from Human Adipose Tissue-Derived Mesenchymal Stem Cells

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Abstract

Understanding of β cell regeneration is needed to develop new treatment modalities in diabetes mellitus. We present our experience of glucose-sensitive insulin-secreting mesenchymal stem cells (IS-MSC) generated and differentiated from human adipose tissue (h-AD) with application of specific differentiation media, sans xenogenic material. h-AD from donor abdominal wall was collected in proliferation medium composed of α-Minimum Essential Media, albumin, fibroblast-growth factor and antibiotics, minced, incubated in collagenase I at 37 °C with shaker and centrifuged. Supernatant and pellets were separately cultured in proliferation medium on cell + plates at 37 °C with 5 % CO2 for 10 days. Cells were harvested, checked for viability, sterility, quantification, flow-cytometry (CD45−/90+/73+), and differentiated into insulin-expressing cells using medium composed of Dulbecco’s modified eagle’s medium, gene expressing upregulators and antibiotics for 3 days. They were studied for transcriptional factors paired box genes-6(Pax-6), islet 1 transcriptional factor (Isl-1), pancreatic and duodenal homobox-1(Pdx-1). C-peptide and insulin were measured by chemiluminescence. IS-MSC showed presence of all three transcriptional factors and showed rise in insulin and c-peptide level in presence of glucose stimuli. It can be concluded that the specific extrinsic factors used in the defined differentiation media effectively and safely promote differentiation of glucose-sensitive insulin-secreting cells from human adipose tissue, without any genetic modulation.

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Abbreviations

ACT A:

Activin A

α-MEM:

Alpha modified Minimum Essential Media

B-hFGF:

Basic-human fibroblast growth factor

DMEM:

Dulbecco’s modified eagle’s medium

EB:

Embryoid bodies

ESC:

Embryonic stem cell

GLP:

Glucagon-like peptide

H-AD-MSC:

Human adipose tissue-derived mesenchymal stem cells

HGF:

Hepatocyte growth factor

Isl-1:

Islet 1 transcriptional factor

ICC:

Islet-like cell clusters

IS-MSC:

Insulin-secreting mesenchymal stem cells

NIC:

Nicotinamide

Pax-6:

Paired box genes-6

PBS:

Phosphate-buffered saline

Pdx-1:

Pancreatic and duodenal homobox 1

Png:

Pentagastrin

RPM:

Revolutions per minute

TGF-β:

Transforming growth factor-β

References

  1. Shapiro, A. M. J., Lakey, J. R. T., Ryan, E. A., Korbutt, G. S., Toth, E., Warnock, G. L., et al. (2000). The New England Journal of Medicine, 343(4), 230–238.

    Article  CAS  Google Scholar 

  2. Frouda, T., Ricordia, C., Baidala Hafiz, M. M., Ponte, G., Cure, P., Pileggi, A., et al. (2005). American Journal of Transplantation, 5(8), 2037–2046.

    Article  Google Scholar 

  3. Bonner-Weir, S., Taneja, M., Weir, G. C., Tatarkiewicz, K., Song, K. H., Sharma, A., et al. (2000). Proceeding National Academy Science USA, 97, 7999–8004.

    Article  CAS  Google Scholar 

  4. Schmied, B. M., Ulrich, A., Matsuzaki, H., Ding, X., Ricordi, C., Weide, L., et al. (2001). Pancreas, 23(2), 157–171.

    Article  CAS  Google Scholar 

  5. Abraham, E. J., Leech, C. A., Lin, J. C., Zulewski, H., & Habener, J. F. (2002). Endocrinology, 143, 3152–3161.

    Article  CAS  Google Scholar 

  6. Lipsett, M., & Finegood, D. T. (2002). Diabetes, 51, 1834–1841.

    Article  CAS  Google Scholar 

  7. Gao, R., Ustinov, J., Pulkkinen, M. A., Lundin, K., Korsgren, O., & Otonkoski, T. (2003). Diabetes, 52(8), 2007–2015.

    Article  CAS  Google Scholar 

  8. Hardikar, A. A., Marcus-Samuels, B., Geras-Raaka, E., Raaka, B. M., & Gershengorn, M. C. (2003). Proceeding National Academy Science USA, 100, 7117–7122.

    Article  CAS  Google Scholar 

  9. Hori, Y., Rulifson, I. C., Tsai, B. C., Heit, J. J., Cahoy, J. D., & Kim, S. K. (2002). Proceeding National Academy Science USA, 99, 16105–16110.

    Article  CAS  Google Scholar 

  10. Kim, D., et al. (2003). Pancreas, 27, E34–E41.

    Article  Google Scholar 

  11. Timper, K., Sebok, D., Eberhardt, M., Linscheid, P., Christ-Crain, M., Keller, U., et al. (2006). Science, 341(4), 1135–1140.

    CAS  Google Scholar 

  12. Sandler, S., Jansson, L., & Welsh, N. (1992). Molecular and Cellular Endocrinology, 86(3), 149–156.

    Article  CAS  Google Scholar 

  13. Del Zotto, H., Massa, L., Rafaeloff, R., Pittenger, G. L., Vinik, A., Gold, G., et al. (2000). Journal of Endocrinology, 165(3), 725–733.

    Article  Google Scholar 

  14. Liu, Y., Song, Z., Zhao, Y., Qin, H., Cai, J., Zhang, H., et al. (2006). Biochemical and Biophysical Research Communications, 346(1), 131–139.

    Article  CAS  Google Scholar 

  15. Uchigata, Y., Yamamoto, H., Nagai, H., & Okamoto, H. (1983). Diabetes, 32, 316–318.

    Article  CAS  Google Scholar 

  16. Yonemura, Y., Takashima, T., Miwa, K., Miyazaki, I., Yamamoto, H., & Okamoto, H. (1984). Diabetes, 33(4), 401–404.

    Article  CAS  Google Scholar 

  17. Sugiyama, K., Yonemura, Y., & Okamoto, H. (1991). International Journal of Pancreatology, 8, 85–95.

    CAS  Google Scholar 

  18. Sandler, S., & Andersson, A. (1986). Diabetologia, 29, 199–202.

    Article  CAS  Google Scholar 

  19. Sandler, S., & Andersson, A. (1988). Transplantation, 46, 30–31.

    Article  CAS  Google Scholar 

  20. Sandler, S., et al. (1989). Diabetes, 38(Suppl.1), 168–171.

    CAS  Google Scholar 

  21. Li, L., Yi, Z., Seno, M., & Kojima, I. (2004). Diabetes, 53(3), 608–615.

    Article  CAS  Google Scholar 

  22. Otonkoski, T., Cirulli, V., Beattie, M., Mally, M. I., Soto, G., Rubin, J. S., et al. (1996). Endocrinology, 137(7), 3131–3139.

    Article  CAS  Google Scholar 

  23. Gahr, S., Merger, M., Bollheimer, L. C., Hammerschmied, C. G., Schölmerich, J., & Hügl, S. R. (2002). Journal of Molecular Endocrinology, 28(2), 99–110.

    Article  CAS  Google Scholar 

  24. Nakano, M., Yasunami, Y., Maki, T., Kodama, S., Ikehara, Y., Nakamura, T., et al. (2000). Transplantation, 69(2), 214–221.

    Article  CAS  Google Scholar 

  25. Garcia-Ocana, A., Vasavada, R. C., Cebrian, A., Reddy, V., Takane, K. K., López-Talavera, J. C., et al. (2001). Diabetes, 50(12), 2752–2762.

    Article  CAS  Google Scholar 

  26. Mashima, H., Yamada, S., Tajima, T., Seno, M., Yamada, H., Takeda, J., et al. (1999). Diabetes, 48(2), 304–309.

    Article  CAS  Google Scholar 

  27. Rooman, I., Lardon, J., & Bouwens, L. (2002). Diabetes, 51, 686–690.

    Article  CAS  Google Scholar 

  28. Bai, L., Meredith, G., & Tuch, B. E. (2005). Journal Endocrinology, 186, 343–352.

    Article  CAS  Google Scholar 

  29. Vanikar, A.V., Dave, S.D., Thakkar, U.G., Trivedi, H.L. (2010) Stem Cells Int. 2010 Dec 20,582382.

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Acknowledgments

The authors are thankful to Ms. Sengunthar Shobhanarani.for statistical analysis, C. N. Patel, J. V. Patel, A. G. Bhargava, and P. N. Bhavsar for carrying out all the laboratory tests including flow-cytometry analysis and chemiluminescence assays and librarian Ms. Jyotsna Suthar for literature survey.

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Authors and Affiliations

  1. Department of Pathology, Laboratory Medicine, Transfusion Services and Immunohematology, G. R. Doshi and K. M. Mehta Institute Of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), Civil Hospital Campus, Asarwa, Ahmedabad, 380016, Gujarat, India

    S. D. Dave & A. V. Vanikar

  2. Department of Nephrology and Transplantation Medicine, G. R. Doshi and K. M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)–Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), Civil Hospital Campus, Asarwa, Ahmedabad, 380016, Gujarat, India

    H. L. Trivedi

  3. Stem Cell Lab, Transplantation Biology Research Centre, Department of Pathology, Laboratory Medicine, Transfusion Services and Immunohematology, G. R. Doshi and K. M. Mehta Institute of Kidney Diseases & Research Centre (IKDRC)- Dr. H.L. Trivedi Institute of Transplantation Sciences (ITS), Civil Hospital Campus, Asarwa, Ahmedabad, 380016, Gujarat, India

    S. D. Dave

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  1. S. D. Dave
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Correspondence to S. D. Dave.

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Dave, S.D., Vanikar, A.V. & Trivedi, H.L. Extrinsic Factors Promoting In Vitro Differentiation of Insulin-Secreting Cells from Human Adipose Tissue-Derived Mesenchymal Stem Cells. Appl Biochem Biotechnol 170, 962–971 (2013). https://doi.org/10.1007/s12010-013-0250-y

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