Directed differentiation of human iPSC into insulin producing cells is improved by induced expression of PDX1 and NKX6.1 factors in IPC progenitors

Journal of Translational Medicine

Research

Anna M. Drozd¹  Maciej P. Walczak¹  Dawid P. Grzela¹  Ewelina Stoczynska-Fidelus²  Piotr Rieske³ 

[1] Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland;Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland;Department of Tumor Biology, Medical University of Łódź, Żeligowskiego 7/9, 90-752, Łódź, Poland;Department of Research and Development, Celther Polska Ltd., Milionowa 23, 93-193, Łódź, Poland;Department of Tumor Biology, Medical University of Łódź, Żeligowskiego 7/9, 90-752, Łódź, Poland;Research and Development Unit, Personather Ltd., Milionowa 23, 93-193, Łódź, Poland;

关键词: Defined culture conditions;    Diabetes;    Differentiation;    Induced pluripotent stem cells;    Insulin producing cells;    NKX6.1;    PDX1;    Reprogramming;   

DOI  :  10.1186/s12967-016-1097-0

 received in 2016-05-25, accepted in 2016-11-24,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundInduced pluripotent stem cells (iPSC) possess an enormous potential as both, scientific and therapeutic tools. Their application in the regenerative medicine provides new treatment opportunities for numerous diseases, including type 1 diabetes. In this work we aimed to derive insulin producing cells (IPC) from iPS cells established in defined conditions.MethodsWe optimized iPSC generation protocol and created pluripotent cell lines with stably integrated PDX1 and NKX6.1 transgenes under the transcriptional control of doxycycline-inducible promoter. These cells were differentiated using small chemical molecules and recombinant Activin A in the sequential process through the definitive endoderm, pancreatic progenitor cells and insulin producing cells. Efficiency of the procedure was assessed by quantitative gene expression measurements, immunocytochemical stainings and functional assays for insulin secretion.ResultsGenerated cells displayed molecular markers characteristic for respective steps of the differentiation. The obtained IPC secreted insulin and produced C-peptide with significantly higher hormone release level in case of the combined expression of PDX1 and NKX6.1 induced at the last stage of the differentiation.ConclusionsEfficiency of differentiation of iPSC to IPC can be increased by concurrent expression of PDX1 and NKX6.1 during progenitor cells maturation. Protocols established in our study allow for iPSC generation and derivation of IPC in chemically defined conditions free from animal-derived components, which is of the utmost importance in the light of their prospective applications in the field of regenerative medicine.

【 授权许可】

CC BY   
© The Author(s) 2016