Growth state and adenovirus infection efficiency of human umbilical cord-derived mesenchymal stem cells in 3 different culture systems
10.3969/j.issn.1673-8225.2010.01.010
- VernacularTitle:人脐带间充质干细胞在3种不同培养体系中的生长状况及腺病毒感染效率
- Author:
Jingxin HONG
;
Qianzhen ZHANG
;
Junling HAN
;
Hui LIU
;
Jian LIU
;
Lugui QIU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2010;14(1):42-47
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: In vitro culture condition and culture efficiency are different in reported umbilical cord-derived mesenchymal stem cells, and lacked of unified standards. Different derived mesenchymal stem cells have different biological properties. Therefore, it is very necessary to establish a simple and high-performance culture system for umbilical cord-derived mesenchymal stem cells. OBJECTIVE: To observe the growth state of human umbilical cord-derived mesenchymal stem cells in different culture systems in vitro and adenovirus infection efficiency. METHODS: Mesenchymal stem cells were separated from healthy full-termed delivery fetus using collagenase digestion method and purified by adherent culture. These cells were cultured and amplified in DMEM (low glucose), MesenPRO RS~(TM) Medium and STEMPRO~(R) MSC SFM in vitro. The 3-5 passage mesenchymal stem cells were infected by the Ad5-EGFP, Ad5/11-EGFP, Ad5/35-EGFP as multiplicity of infection (MOI)=1, 10, 100. Viral infection and green fluorescence expression were observed at post-infection 24, 56 and 72 hours using inverted fluorescence microscope. RESULTS AND CONCLUSION: The cell morphology in STEMPR~(R) MSC SFM was different from other two culture system and these cells were not easy to adherent after trypsin digestion. Cell doubling time in the MesenPRO RS~(TM) Medium was shorter than other two groups. Mesenchymal stem cells were infected by Ad5/35-EGFP with higher efficiency than other two kinds of adenovirus, but part of cells appeared apoptosis. The infection efficiency of Ad5/11-EGFP was highest. The fluorescence intensity was gradually increased with increased MOI.
