Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (19): 2988-2993.doi: 10.3969/j.issn.2095-4344.3522

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Comparison of biological characteristics of adipose derived mesenchymal stem cells in Wuzhishan inbreed miniature pigs aged two different months

Gao Yuanhui1, Xiang Yang2, Cao Hui1, Wang Shunlan1, Zheng Linlin1, He Haowei1, Zhang Yingai1, Zhang Shufang1, Huang Denggao1   

  1. 1Central Laboratory, 2Department of Urology, Affiliated Haikou Hospital of Xiangya School of Medicine, Central South University, Haikou 570208, Hainan Province, China
  • Received:2020-05-19 Revised:2020-05-20 Accepted:2020-06-29 Online:2021-07-09 Published:2021-01-13
  • Contact: Huang Denggao, Master, Assistant researcher, Central Laboratory, Affiliated Haikou Hospital of Xiangya School of Medicine, Central South University, Haikou 570208, Hainan Province, China
  • About author:Gao Yuanhui, Master candidate, Central Laboratory, Affiliated Haikou Hospital of Xiangya School of Medicine, Central South University, Haikou 570208, Hainan Province, China
  • Supported by:
    the Natural Youth Fund of Hainan Province, No. 819QN390 (to GYH); the Natural Youth Fund of Hainan Province, No. 819QN386 (to HDG); the Major Science and Technology Plan Project of Hainan Province, No. ZDKJ2017007 (to ZSF)

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Abstract: BACKGROUND: Stem cells have been the focus of regenerative medicine research. Many factors affect the biological characteristics of stem cells, among which donor age is one of the critical factors.
OBJECTIVE: To observe biological characteristics of two types of inbreed line miniature pigs of Wuzhishan adipose-derived mesenchymal stem cells so as to screen the optimal seed cell source.
METHODS:  Inbreed line miniature pigs of Wuzhishan adipose-derived mesenchymal stem cells aged 4 and 12 months were selected as materials. The passage 3 and passage 10 of cells were selected to observe their cell morphology, draw cell proliferation curve, and detect cell surface antigen, cell cycle and apoptosis. Afterwards, osteogenic differentiation was performed with Alizarin red staining, followed by adipogenic differentiation with Oil Red O staining and cell senescence β-galactosidase staining.
RESULTS AND CONCLUSION: (1) The morphology of cells at 4 and 12 months both showed vortex-shaped adherent growth. The expression of surface markers CD29, CD44 and CD90 was positive, while that of CD8a, CD45 and HLA-DR was negative. (2) The proliferation of cells at 4 and 12 months was stronger in passage 3. In passage 10, the percentage of cells in the G0/G1 phase (P < 0.001) and early apoptosis (P < 0.001) increased, while in the S and G2/M phases, it decreased at 12 months compared with 4 months (P < 0.001). (3) There was no significant difference in the differentiation ability of cells at passage 3 in vitro between 4 and 12 months. The calcification nodule rate and lipid droplet rate of cells at 12 months of passage 10 were significantly lower than at 4 months of passage 10 (P < 0.001). (4) The β-galactosidase of cells was almost not expressed at 4 and 12 months of passage 3. However, in the passage 10, the positive rate of cells at 12 months was significantly higher than that at 4 months (P < 0.001); and the cell morphology became more flattened, hypertrophic and less refractive. (5) It is concluded that adipose-derived mesenchymal stem cells of inbreed line miniature pigs of Wuzhishan at 4 months showed stronger proliferation and differentiation ability, while showed lower senescence on β-galactosidase activity, suggesting that 4-month cells are more suitable for autograft. 

Key words: stem cells, adipose mesenchymal stem cells, inbred Wuzhishan miniature pig, age, proliferation, differentiation, senescence

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