Open AccessIn vitro quantitative and relative gene expression analysis of pancreatic transcription factors P dx‐1, N gn‐3, I sl‐1, P ax‐4, P ax‐6 and N kx‐6.1 in trans‐differentiated human hepatic progenitors
Author(s) -
Vishwakarma Sandeep Kumar,
Rahamathulla Syed,
Bardia Avinash,
Tiwari Santosh K,
Srinivas Gunda,
Raj Avinash,
Tripura Chaturvedula,
Sandhya Annamaneni,
Habeeb Mohammed Aejaz,
Khan Aleem A,
Pande Gopal,
Reddy K Pratap,
Reddy P Yugandhar
Publication year - 2014
Publication title -
journal of diabetes investigation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.089
H-Index - 50
eISSN - 2040-1124
pISSN - 2040-1116
DOI - 10.1111/jdi.12193
Subject(s) - progenitor cell , medicine , cell , in vitro , transcription factor , diabetes mellitus , gene expression , endocrinology , andrology , microbiology and biotechnology , biology , gene , stem cell , genetics
Aims/Introduction Diabetes is a major health concern throughout the world because of its increasing prevalence in epidemic proportions. β‐Cell deterioration in the pancreas is a crucial factor for the progression of diabetes mellitus. Therefore, the restoration of β‐cell mass and its function is of vital importance for the development of effective therapeutic strategies and most accessible cell sources for the treatment of diabetes mellitus. Materials and Methods Human fetuses (12–20 weeks gestation age) were used to isolate human hepatic progenitor cells ( hHPC s) from fetal liver using a two‐step collagenase digestion method. Epithelial cell adhesion molecule‐positive (EpCAM+ve)‐enriched hHPC s were cultured in vitro and induced with 5–30 mmol/L concentration of glucose for 0–32 h. Pdx‐1 expression and insulin secretion was analyzed using immunophenotypic and chemifluorescence assays, respectively. Relative gene expression was quantified in induced hHPC s, and compared with uninduced and pancreatic cells to identify the activated transcription factors (Pdx‐1, Ngn‐3, Isl‐1, Pax‐4, Pax‐6 and Nkx‐6.1) involved in β‐cell production. Results EpCAM+ve cells derived from human fetal liver showed high in vitro trans‐differentiation potential towards the β‐cell phenotype with 23 mmol/L glucose induction after 24 h. The transcription factors showed eminent expression in induced cells. The expression level of transcription factors was found significantly high in 23 mmol/L‐induced hHPC s as compared with the uninduced cells. Conclusions The present study has shown an exciting new insight into β‐cell development from hHPC s trans‐differentiation. Relative quantification of gene expression in trans‐differentiated cells offers vast possibility for the production of a maximum number of functionally active pancreatic β‐cells for a future cure of diabetes.