title: In vitro quantitative and relative gene expression analysis of pancreatic transcription factors Pdx-1, Ngn-3, Isl-1, Pax-4, Pax-6 and Nkx-6.1 in trans-differentiated human hepatic progenitors.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  Human fetuses (12-20 weeks gestation age) were used to isolate human hepatic progenitor cells (hHPCs) from fetal liver using a two-step collagenase digestion method. Epithelial cell adhesion molecule-positive (EpCAM+ve)-enriched hHPCs 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 hHPCs, 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 hHPCs as compared with the uninduced cells.
  
  
  CONCLUSIONS
  The present study has shown an exciting new insight into β-cell development from hHPCs 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.
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Sandeep Kumar
authors.initials: SK
authors.lastname: Vishwakarma
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Syed
authors.initials: S
authors.lastname: Rahamathulla
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Avinash
authors.initials: A
authors.lastname: Bardia
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Santosh K
authors.initials: SK
authors.lastname: Tiwari
authors.affiliation: Center for Cellular and Molecular Biology (CCMB) Osmania University Hyderabad Andhra Pradesh India.
authors.forename: Gunda
authors.initials: G
authors.lastname: Srinivas
authors.affiliation: Center for Cellular and Molecular Biology (CCMB) Osmania University Hyderabad Andhra Pradesh India.
authors.forename: Avinash
authors.initials: A
authors.lastname: Raj
authors.affiliation: Center for Cellular and Molecular Biology (CCMB) Osmania University Hyderabad Andhra Pradesh India.
authors.forename: Chaturvedula
authors.initials: C
authors.lastname: Tripura
authors.affiliation: Department of Genetics Osmania University Hyderabad Andhra Pradesh India.
authors.forename: Annamaneni
authors.initials: A
authors.lastname: Sandhya
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Mohammed Aejaz
authors.initials: MA
authors.lastname: Habeeb
authors.affiliation: Center for Liver Research and Diagnostics (CLRD) Deccan College of Medical Sciences Kanchanbagh Hyderabad Andhra Pradesh India.
authors.forename: Aleem A
authors.initials: AA
authors.lastname: Khan
authors.affiliation: Center for Cellular and Molecular Biology (CCMB) Osmania University Hyderabad Andhra Pradesh India.
authors.forename: Gopal
authors.initials: G
authors.lastname: Pande
authors.affiliation: Department of Zoology Osmania University Hyderabad Andhra Pradesh India.
authors.forename: K Pratap
authors.initials: KP
authors.lastname: Reddy
authors.affiliation: Department of Zoology The Adony Arts and Science College Kurnool India.
authors.forename: P Yugandhar
authors.initials: PY
authors.lastname: Reddy
keywords: Pancreatic transcription factors; Relative quantification; Trans‐differentiation