Generation of Induced Hepatocyte-Like Cells from Fibroblasts by Defined Factors

Royan International Twin Congress 12th Congress on Stem Cell Biology and Technology

31 August till 2 September 2016، Tehran - Iran

Presentation Type: Speech

Objective: The expandable patient–specific induce hepatocyte- like cells (iHLCs) that are generated through direct conversion can elucidate tumorigenic potential that induced pluripotent stem cells (hiPSCs) has. Thus, the generation of iHLCs has a potential to be used as a cell source for regenerative medicine and drug screening in liver disease. Materials and Methods: To generated iHLCs, defined transcription factors were delivered to fibroblasts by lentivirus. The cell fate converting process was examined by observing the morphological changes. The expression of hepatic genes and proteins was examined by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence staining. To evaluate the functions of iHLCs in vitro, low-density lipoprotein (LDL) uptake assay, indocyanine green (ICG) test, periodic acid-schiff stain, and enzyme-linked immunosorbent assay (ELISA) were conducted. Results: iHLCs were generated via direct conversion using define transcription factors in combination with lineage specific culture condition. iHLCs showed typical morphology of hepatocytes, hepatic markers and functions which include glycogen storage, LDL uptake, ICG metabolism, and Albumin secretion. Conclusion: This study demonstrates a novel set of transcription factors in converting fibroblasts into functional hepatocytes which would be used in therapeutic applications.