BM-MSCs Differentiated Insulin-Producing Cells Produce More Insulin in Presence of EGF than of FGF

Cheia Majeed¹, Ahmed M. Al-Shammari^2*, Eman H. Yausif¹, Nahi Y. Yaseen², Ayman Ali Husien², Rajha A. Al-Naimi¹, Adnan Ajeel²

¹Department of Pathology, College of Veterinary Medicine, Baghdad University, Baghdad, Iraq.
²Experimental Therapy Department, Iraqi Center for Cancer and Medical Genetic Research, Al-Mustansiriya University, Baghdad, Iraq.

DOI: 10.4236/scd.2015.54004                

Abstract

Adult stem cells have the ability to differentiate into islet like cells for the hope of treating diabetes mellitus (DM). The most important part is the differentiation process from na?ve stem cells to fully differentiated fully functional islet cells. For this purpose, we wanted to optimize the current proposed differentiation media by replacing the FGF with EGF and measure insulin production. Bone marrow-derived MSCs were from mice long bones and expanded in cell culture before induction of differentiation. Stem cells surface markers were analyzed by immunocytochemistry. Cultured stem cells were negative for CD34 while they expressed high levels of CD90. Differentiated cells morphology was studied by using H & E stain. Differentiated cells were detected by studying protein expression of insulin as specific marker for IPC differentiation. Cells function was studied by measuring the insulin production in tissue culture supernatant in vitro and also insulin release in response to glucose challenge. Ditizone staining were both positive. Insulin was secreted by these cells in response to different concentrations of glucose stimulation in a regulated manner. Cells induced with formula contain EGF produced more insulin in the same formula but contain FGF instead, this prove that EGF is the best to use during differentiation process.

Keywords

Diabetes Mellitus, Pancreatic β Cells, Growth Factors

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Zhang, Y.H., Shen, W.Z., Hua, J.L., Lei, A.M., Lv, C.R., Wang, H.Y., Yang, C.R., Gao, Z.M. and Dou, Z.Y. (2010) Pancreatic Islet-Like Clusters from Bone Marrow Mesenchymal Stem Cells of Human First-Trimester Abortus Can Cure Streptozocin-Induced Mouse Diabetes. Rejuvenation Research, 13, 695-706. http://dx.doi.org/10.1089/rej.2009.1016
[2]	Raju, S.M. and Raju, B. (2010) Illustrated Medical Biochemistry. 2nd Edition, Jaypee Brothers Medical Publishers Ltd., New Delhi.
[3]	Kang, H.M., Kim, J., Park, S., Kim, J., Kim, H., Kim, K.S., et al. (2009) Insulin-Secreting Cells from Human Eyelid-Derived Stem Cells Alleviate Type I Diabetes in Immunecompetent Mice. Stem Cells, 27, 1999-2008.
[4]	Morisot, C.F., Fraipont, D.E., Richard, M., Marchand, M., Savatier, P., Bosco, D., Favrot, M. and Benhamou, P. (2005) Human Bone Marrow Mesenchymal Stem Cell Can Express Insulin and Key Transcription Factors of the Endocrine Pancreas Developmental Pathway upon Genetic and/or Micro Environmental Manipulation in Vitro. Stem Cells, 23, 594-603. http://dx.doi.org/10.1634/stemcells.2004-0123
[5]	Gabr, M., Sobh, M., Zakaria, M., Refaie, A. and Ghoneim, M. (2008) Transplantation of Insulin-Producing Clusters Derived from Adult Bone Marrow Stem Cells to Treat Diabetes in Rats. Experimental and Clinical Transplantation, 6, 236-243.
[6]	Taneera, J., Rosengren, A., Renstrom, E., Nygren, J.M., Serup, P., Rorsman, P., et al. (2006) Failure of Transplanted Bone Marrow Cells to Adopt a Pancreatic β-Cells Fate. Diabetes, 55, 290-296. http://dx.doi.org/10.2337/diabetes.55.02.06.db05-1212
[7]	Evans, C.M., Vestermark, G.L. and Mirmira, R.G. (2009) Development of Insulin-Producing Cells from Primitive Biologic Precursors. Current Opinion in Organ Transplantation, 14, 56-63. http://dx.doi.org/10.1097/MOT.0b013e3283186fc1
[8]	Zhang, Y., Li, C.D., Jiang, X.X., Li, H.L., Tang, P.H. and Mao, N. (2004) Comparison of Mesenchymal Stem Cells from Human Placenta and Bone Marrow. Chinese Medical Journal, 117, 882-887.
[9]	Yang, L., Li, S., Hatch, H., Ahrens, K., Cornelius, J.G., Petersen, B.E. and Peck, A.B. (2002) In Vitro Trans-Differentiation of Adult Hepatic Stem Cells into Pancreatic Endocrine Hormone-Producing Cells. Proceedings of the National Academy of Sciences of the United States of America, 99, 8078-8083.
[10]	Guo, Z., Yang, J., Liu, X., Li, X., Hou, C., Tang, P.H., et al. (2001) Biological Features of Mesenchymal Stem Cells from Human Bone Marrow. Chinese Medical Journal, 114, 950-953.
[11]	Kern, S., Eichler, H., Stoeve, J., Klular, H. and Bieback, K. (2006) Comparative Analysis of Mesenchymal Stem Cells from Bone Marrow, Umbilical Cord Blood, or Adipose Tissue. Stem Cell, 24, 1294-1301 http://dx.doi.org/10.1634/stemcells.2005-0342
[12]	Soria, B., Skoudy, A. and Martín, F. (2001) From Stem Cells to Beta Cells: New Strategies in Cell Therapy of Diabetes mellitus. Diabetologia, 44, 407-415.
[13]	D’Amour, K., Bang, A., Eliazer, S., Kelly, O., Aguluick, A., Smart, N., Moorman, M., Kroon, E., Carpenter, M. and Baetge, E. (2006) Production of Pancreatic Hormone-Expressing Endocrine Cells from Human Embryonic Stem Cells. Nature Biotechnology, 24, 1392-1401. http://dx.doi.org/10.1038/nbt1259
[14]	Kim, Y., Lee, J., Shin, J., Kim, H. and Kim, C. (2003) Enhancement of Mouse Pancreatic Regeneration and HIT-T15 Cell Proliferation with Rat Pancreatic Extract. Biochemical and Biophysical Research Communications, 309, 528-532. http://dx.doi.org/10.1016/j.bbrc.2003.07.011
[15]	Cras, M.C., Elghazi, L., Czernichow, P. and Scharfmann, R. (2001) Epidermal Growth Factor Increases Undifferentiated Pancreatic Embryonic Cells in Vitro: A Balance between Proliferation and Differentiation. Diabetes, 50, 1571-1579. http://dx.doi.org/10.2337/diabetes.50.7.1571
[16]	Elghazi, L., Cras, M.C., Czernichow, P. and Scharfmann, R. (2002) Role for FGFR2IIIb-Mediated Signals in Controlling Pancreatic Endocrine Progenitor Cell Proliferation. Proceedings of the National Academy of Sciences of the United States of America, 99, 3884-3889. http://dx.doi.org/10.1073/pnas.062321799
[17]	Kramer, B. and Penny, C. (2001) Regulation of Embryonic Chick Insulin Cells: Effect of Retinoic Acid and Insulin-Like Growth Factor 1. Cells Tissues Organs, 169, 42-48. http://dx.doi.org/10.1159/000047859
[18]	Tang, D.-Q., Cao, L.-Z., Burckhardt, B., Xia, C.-Q., Litherl, S.A., Atkinson, M.A. and Yang, L.-Y. (2004) In Vivo and in Vitro Characterization of Insulin-Producing Cells Obtained from Murine Bone Marrow. Diabetes, 53, 1721-1732. http://dx.doi.org/10.2337/diabetes.53.7.1721
[19]	Xu, G., Stoffers, D., Habener, J. and Bonner-Weir, S. (1999) Exendin-4 Stimulates both Beta-Cell Replication and Neogenesis, Resulting in increased Mass and Improved Glucose Tolerance in Diabetic Rats. Diabetes, 48, 2270-2276. http://dx.doi.org/10.2337/diabetes.48.12.2270
[20]	Bouwens, L. and Rooman, I. (2005) Regulation of Pancreatic Beta-Cell Mass. Physiological Reviews, 85, 1255-1270. http://dx.doi.org/10.1152/physrev.00025.2004