Isolation, culture and biological characteristics of adipose-derived mesenchymal stem cells from patients with hepatitis B virus infection

Wang Yan-hui; Zhang Shu-qin; Zhao Wen-jing

doi:10.3969/j.issn.2095-4344.2015.36.009

Abstract：BACKGROUND:Chronic hepatitis B virus infection can impact the biological characteristics of bone marrow mesenchymal stem cel s. Adipose-derived mesenchymal stem cel s have gained more and more attention due to their high safety, little invasiveness, easy purification and rapid proliferation. OBJECTIVE:To establish the isolation and culture method of adipose-derived mesenchymal stem cel s from patients with hepatitis B virus infection in vitro, and to observe the biological characteristics of cel s. METHODS:Adipose-derived mesenchymal stem cel s were isolated from the subcutaneous fat of hepatitis B virus infection patients by col agenase digestion and adherent method. Growth curve of adipose-derived mesenchymal stem cel s were detected by MTT method and cel phenotypes were detected by flow cytometry. The adipogenic and osteogenic differentiation potential of adipose-derived mesenchymal stem cel s were detected in vitro. RESULTS AND CONCLUSION:(1) Adipose-derived mesenchymal stem cel s from 10 patients with hepatitis B virus infection were al isolated and cultured successful y. The primary passage time of adipose-derived mesenchymal stem cel s was (8.3±1.2) days. The growth curve of cel s was“S”shaped. Cel s came into a logarithmic phase at days 3-4, and came into platform at day 7. (3) Passage 3 adipose-derived mesenchymal stem cel s highly expressed CD29, CD166, HLA-ABC and CD44, but did not express or lowly expressed CD34 and HLA-DR. (3) Adipose-derived mesenchymal stem cel s differentiated into adipocytes after adipogenic induction, and differentiated cel s were positive for oil red O staining;after osteogenic induction, adipose-derived mesenchymal stem cel s differentiated into osteoblasts that were positive for alkaline phosphatase staining. These findings indicate that the col agenase digestion and adherent method can be used to effectively isolate adipose-derived mesenchymal stem cel s from patients with hepatitis B virus infection, and the cel proliferation is rapid so that a large number of cel s can be obtained in the short term.