Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (15): 2318-2324.doi: 10.12307/2023.392

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Biological characteristics of human amniotic mesenchymal stem cells pretreated with different volume fractions of oxygen

Tang Jianhong1, Zhang Nini2, Huang Guilin2, Long Yuanzhu2, Cui Tianning1, Luo Qinliang1, Lang Jiachan1, Dai Min2, Zhang Ligang2   

  1. 1Zunyi Medical University, Zunyi 563000, Guizhou Province, China; 2Department of Oral and Maxillofacial Surgery, Affiliated Stomatological Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, China
  • Received:2022-05-20 Accepted:2022-07-14 Online:2023-05-28 Published:2022-10-17
  • Contact: Zhang Nini, Master, Associate professor, Department of Oral and Maxillofacial Surgery, Affiliated Stomatological Hospital of Zunyi Medical University, Zunyi 563000, Guizhou Province, China
  • About author:Tang Jianhong, Master, Physician, Zunyi Medical University, Zunyi 563000, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81860198 (to ZNN); National Natural Science Foundation of China, No. 81960204 (to HGL); Master’s Start-up Fund of Zunyi Medical University, No. KY2019-1 (to DM); a grant from Guizhou Provincial Health Commission, No. gzwjkj2020-1-165 (to ZLG); “Future Clinical Famous Doctor” Project of Zunyi Medical University, No. 20211017 (to ZNN)

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Abstract: BACKGROUND: In recent years, stem cell plays an important role in the repair of tissue injury, among which mesenchymal stem cell transplantation is the most widely used. Mesenchymal stem cells can secrete cytokines through paracrine effect to promote the repair of damaged tissue. Hypoxia pretreatment can improve the biological characteristics of mesenchymal stem cells. Hypoxic preconditioning of human amniotic mesenchymal stem cells may play a better therapeutic role in tissue injury repair. 
OBJECTIVE: To study the effects of different hypoxic pretreatment on the proliferation, antiapoptotic ability, and parocrine effect of human amniotic mesenchymal stem cells.  
METHODS: The primary human amniotic mesenchymal stem cells were obtained by enzyme digestion method. Passage 3 human amniotic mesenchymal stem cells were pretreated with different hypoxic concentrations for 48 hours and randomly divided into hypoxic pretreatment group (1% O2, 3% O2, 5% O2, and 10% O2) and control group (21% O2). The morphology of human amniotic mesenchymal stem cells was observed under an inverted microscope. The proliferation ability of human amniotic mesenchymal stem cells was detected by CCK-8 assay. Annexin V-FITC/PI kit was used to detect the apoptosis of human amniotic mesenchymal stem cells. The apoptosis gene and hypoxia-inducible factor of human amniotic mesenchymal stem cells were detected by real-time quantitative PCR. The cytokine secretion capacity of human amniotic mesenchymal stem cells in each group was detected by enzyme-linked immunosorbent assay.  
RESULTS AND CONCLUSION: Under the microscope, the morphology of human amniotic mesenchymal stem cells in different hypoxic pretreatment groups showed no significant difference. Hypoxic pretreatment could enhance the proliferation, anti-apoptotic and paracrine cytokine capacity of human amniotic mesenchymal stem cells. Pretreatment effect of 1% O2 concentration was better than that of 3% O2, 5% O2, 10% O2 and 21% O2.   

Key words: human amniotic mesenchymal stem cell, hypoxia, pretreatment, biological characteristics

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