Conversion of immortal liver progenitor cells into pancreatic endocrine progenitor cells by persistent expression of Pdx-1.
The conversion of expandable liver progenitor cells into pancreatic beta cells would provide a renewable cell source for diabetes cell therapy. Previously, we reported the establishment of liver epithelial progenitor cells (LEPCs). In this work, LEPCs were modified into EGFP/Pdx-1 LEPCs, cells with stable expression of both Pdx-1 and EGFP. Unlike previous work, with persistent expression of Pdx-1, EGFP/Pdx-1 LEPCs acquired the phenotype of pancreatic endocrine progenitor cells rather than giving rise to insulin-producing cells directly. EGFP/Pdx-1 LEPCs proliferated vigorously and expressed the crucial transcription factors involved in beta cell development, including Ngn3, NeuroD, Nkx2.2, Nkx6.1, Pax4, Pax6, Isl1, MafA and endogenous Pdx-1, but did not secrete insulin. When cultured in high glucose/low serum medium supplemented with cytokines, EGFP/Pdx-1 LEPCs stopped proliferating and gave rise to functional beta cells without any evidence of exocrine or other islet cell lineage differentiation. When transplanted into diabetic SCID mice, EGFP/Pdx-1 LEPCs ameliorated hyperglycemia by secreting insulin in a glucose regulated manner. Considering the limited availability of beta cells, we propose that our experiments will provide a framework for utilizing the immortal liver progenitor cells as a renewable cell source for the generation of functional pancreatic beta cells.
Jin
Cai-Xia
CX
Department of Cell Biology, Second Military Medical University, Shanghai 200433, P. R. China.
Li
Wen-Lin
WL
Xu
Fang
F
Geng
Zhen H
ZH
He
Zhi-Ying
ZY
Su
Juan
J
Tao
Xin-Rong
XR
Ding
Xiao-Yan
XY
Wang
Xin
X
Hu
Yi-Ping
YP
D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D017690:Cell Transplantation; D004847:Epithelial Cells; D022781:Hepatocytes / Q000166:cytology*; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D006943:Hyperglycemia / Q000628:therapy; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000637:transplantation; D051379:Mice; D016513:Mice, SCID; D013234:Stem Cells / Q000166:cytology*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D014157:Transcription Factors / Q000096:biosynthesis
D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D007328:Insulin; C000715627:Nkx2-2 protein, mouse; D015534:Trans-Activators; D014157:Transcription Factors; C084010:pancreatic and duodenal homeobox 1 protein