Role of BMP Signaling in Pancreatic Progenitor Differentiation from Human Embryonic Stem Cells
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Transplantation of pancreatic progenitors derived from human embryonic stem cells (hESCs) is a promising way to treat diabetes. Strategies to obtain the required cell mass would rely on the up scaling of current differentiation protocols, or the proliferation of committed progenitors. We aimed at finding conditions that maintain a proliferating pancreatic progenitor pool and we assessed the role of BMP4 signaling in this process. hESCs were differentiated into PDX1 positive pancreatic progenitor stage following our established protocol with few modifications, and then the progenitor cells were passaged in a defined proliferation medium (PM). During passage, the effect of BMP4 signaling on the differentiation and proliferation of pancreatic progenitors was examined by RT-PCR and immunofluorescence analysis. We found that PDX1 positive pancreatic progenitors proliferated and gained NKX6.1 expression in the PM, whereas they failed to express NKX6.1 if BMP signaling was inhibited with Noggin. In this latter condition, part of the progenitors rather generated pro-endocrine cells denoted by NGN3 and synaptophysin expression. On the contrary, addition of BMP4 to the PM promoted the early derivation of PDX1 and NKX6.1 coexpressing pancreatic progenitors. Our findings are in line with mouse pancreas development, and indicate that BMP4 signaling is required for the derivation and maintenance of hESC-derived PDX1+NKX6.1+ pancreatic progenitors. These results are instructive for guiding the development of an efficient pancreas differentiation protocol in view of diabetes cell replacement therapy.
KeywordshESCs Pancreatic progenitors Proliferation BMP4
PDX1 positive pancreatic progenitor
PDX1 or NKX6.1 positive pancreatic progenitor
PDX1 and NKX6.1 coexpressing pancreatic progenitor
The authors thank E. De Blay, M. Baekeland and W. Rabiot for technical assistance. They are also grateful to A.K. Baczmanska and K. Dee for the maintenance of undifferentiated hESCs cultures. L. Sui is a recipient of the CSC scholarship (China). J.K. Mfopou is a postdoc fellow of the VUB. This work was supported by a Vrije Universiteit Brussel (VUB) research grant under the code GOA41.
LS designed and conducted the study, analyzed the results, wrote and approved the final manuscript. MG and KS provided the hESC line and approved the final manuscript. LB contributed to study design, reviewed and discussed the data, critically reviewed and approved the final manuscript. JKM designed the study, reviewed and discussed the data, wrote and approved the final manuscript.
Conflict of interest
The authors declare no potential conflicts of interest associated with this manuscript.
- 7.Matveyenko, A. V., Georgia, S., Bhushan, A., & Butler, P. C. (2010). Inconsistent formation and nonfunction of insulin-positive cells from pancreatic endoderm derived from human embryonic stem cells in athymic nude rats. American Journal of Physiology. Endocrinology and Metabolism, 299, E713–E720.PubMedCrossRefGoogle Scholar
- 10.Sui, L., Mfopou, J. K., Chen, B., Sermon, K., Bouwens, L. (2012). Transplantation of human embryonic stem cell-derived pancreatic endoderm reveals a site-specific survival, growth and differentiation. Cell Transplantation. doi: 10.3727/096368912X636812.
- 13.Sui, L., Mfopou, J. K., Geens, M., Sermon, K., Bouwens, L. (2012). FGF signaling via MAPK is required early and improves Activin A-induced definitive endoderm formation from human embryonic stem cells. Biochemical and Biophysical Research Communications, 426(3):380–385.Google Scholar
- 16.Thatava, T., Nelson, T. J., Edukulla, R., et al. (2011). Indolactam V/GLP-1-mediated differentiation of human iPS cells into glucose-responsive insulin-secreting progeny. Gene Therapy, 18, 283–293.Google Scholar
- 19.Kobberup, S., Schmerr, M., Dang, M. L., et al. (2010). Conditional control of the differentiation competence of pancreatic endocrine and ductal cells by Fgf10. Mechanisms of Development, 127, 220–234.Google Scholar
- 20.Sneddon, J. B., Borowiak, M., Melton, D. A. (2012). Self-renewal of embryonic-stem-cell-derived progenitors by organ-matched mesenchyme. Nature 491, 765–768.Google Scholar
- 23.Chung, W. S., Andersson, O., Row, R., Kimelman, D., & Stainier, D. Y. (2010). Suppression of Alk8-mediated Bmp signaling cell-autonomously induces pancreatic beta-cells in zebrafish. Proceedings of the National Academy of Sciences of the United States of America, 107, 1142–1147.PubMedCrossRefGoogle Scholar
- 24.Ahnfelt-Ronne, J., Ravassard, P., Pardanaud-Glavieux, C., Scharfmann, R., Serup, P. (2010). Mesenchymal bone morphogenetic protein signaling is required for normal pancreas development. Diabetes, 59, 1948–1956.Google Scholar
- 25.Cai, J., Yu, C., Liu, Y., et al. (2010). Generation of homogeneous PDX1+ pancreatic progenitors from human ES cell-derived endoderm cells. Journal of Molecular Cell Biology, 2, 50–60.Google Scholar