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Updated: May 4, 2026
Isolation and Expansion of Mesenchymal Stem/Stromal Cells Derived from Human Placenta Tissue
Published on: June 6, 2016
[Long term in-vitro expansion reduces immune modulation function of placental chorionic villi mesenchymal stem cells]
Zhou-Xin Yang1, Yue-Ru Ji1, Zhi-Bo Han2
1State Key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
Abstract:
The main aim of this study was to investigate the biological activities and immune modulation changes of chorionic villi mesenchymal stem cells (CV-MSC) after long term culture. The morphology of the CV-MSC of passage 3 and passage 9 were observed by microscopy, and their phenotypes were detected by flow cytometry. CV-MSC of passage 3 and 9 were co-cultured with PHA-stimulated PBMNC, and IFN-γ concentration in culture medium was detected by ELISA. The mRNA expression of COX-2, HGF and HLA-G in CV-MSC were detected by real-time PCR. The results showed that after long term culture, the CV-MSC kept the MSC morphology and most of the phenotypes including CD31, CD34, CD44, CD45, CD62L, CD73, CD90, CD105, CD117, CD151, CD235a, CD271 and HLA-DR, while the CD49d was significantly up-regulated. Immune modulation ability of CV-MSC was reduced and the mRNA expression of COX-2 and HGF was down regulated after long term culture, but the expression of HLA-G did not found to be obvious change. It is concluded that the long term in vitro expansion changes the expression of CD49d and reduces immune modulation of CV-MSC.
Insights
Long term culture of chorionic villi mesenchymal stem cells (CV-MSC) maintains their core characteristics but alters CD49d expression and reduces immune modulation capabilities. These findings are crucial for understanding CV-MSC behavior in regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Immunology
- Regenerative Medicine
Background:
- Chorionic villi mesenchymal stem cells (CV-MSC) are promising for therapeutic applications.
- Understanding the impact of long-term culture on CV-MSC properties is essential for their clinical use.
Purpose of the Study:
- To investigate the biological activities and immune modulation changes of CV-MSC after prolonged in vitro expansion.
- To assess the effects of long-term culture on CV-MSC morphology, phenotype, and immune function.
Main Methods:
- Microscopy and flow cytometry were used to evaluate CV-MSC morphology and phenotype at passage 3 and passage 9.
- Co-culture assays with PHA-stimulated peripheral blood mononuclear cells (PBMNC) and ELISA were performed to measure IFN-γ levels.
- Real-time PCR was employed to detect the mRNA expression of COX-2, HGF, and HLA-G.
Main Results:
- CV-MSC maintained their morphology and most phenotypes after long-term culture, with significant upregulation of CD49d.
- Immune modulatory capacity of CV-MSC decreased, evidenced by reduced IFN-γ levels.
- mRNA expression of COX-2 and HGF was downregulated, while HLA-G expression showed no significant change.
Conclusions:
- Long-term in vitro expansion of CV-MSC leads to altered CD49d expression and diminished immune modulatory functions.
- These changes impact the potential therapeutic applications of CV-MSC, necessitating careful consideration of culture duration.
