Evaluation of the modification of human adipose-derived stem cell induced by hepatic growth factor and platelet-rich-plasma

Abstract

Objectives: The aim of this project is to evaluate functional and safety changes of Human adipose-derived stem cells (hADSCs) under culture condition which Fetal bovine serum replaced by Platelet-rich-plasma (PRP) supplied with Hepatocyte growth factor (HGF), with the hope to find the optimal condition culture medium to culture cells used for cirrhosis transplantation. Method: hADSC cells were thawed and cultured with different kinds of media, including: MSC culture medium, MSC supplied with HGF, MSC serum free supplied with PRP, MSC serum free supplied with both PRP and HGF. After 7 days, cells were evaluated about morphology changing by giemsa staining, cell proliferation by MTT assay, chromosome quality by karyotyping, hepatic enriched protein by immunocytochemistry, hepatic enriched genes by qRT PCR. Results: hADSC after being induced with growth factors still have stem cell morphology and characteristics, proliferation rate are different between control group and treatment groups, they do not express hepatic enriched proteins, such as Alpha anti trypsin I (AAT), Albumin (ALB), Alpha feto protein (AFP) but do express hepatic genes, Cytokeratin 18 (CK18) and Cytokeratin 19 (CK19). Conclusion: this research shows very promising results. Further study can be performed base on this research to find optimal culture condition for cells to be transplanted into mice model or clinically applied as a treatment for cirrhosis patients. Abbreviation hADSCs: Human Adiposed-derived stem cells; PRP: Platelet-rich-plasma; HGF: Hepatocyte growth factor; FCM: Flow cytometry; ICC: Immunocytochemistry; MSCs: Mesenchymal stem cells; ES: Embryonic stem cell; AS: Adult stem cell; FBS: Fetal bovine serum; PBS: Phosphate buffered saline.