Liu Lingying, Chai Jiake, Hou Yusen, Duan Hongjie, Yu Yonghui, Yin Huinan, Hu Quan, Fan Jun, Du Jundong
Department of Burn and Plastic Surgery, First Hospital Affiliated to General Hospital of Chinese PLA, Beijing, 100048, P.R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Jul;27(7):769-74.
To investigate the effect of the serum from severe burn patients on the biology characteristics of human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro, so as to explore the feasibility of hUCMSCs transplantation for treating severe burn.
The 3rd passage of hUCMSCs were randomly divided into 3 groups: 10% fetal bovine serum group (group A), 10% normal serum group (group B), and 10% burn serum group (group C). At 24 hours, 72 hours, and 6 days after culture, the cell morphology and density were observed by inverted microscope; the cell proliferation was assessed by MTT; after 6 days of culture, the cell cycle by propidium iodide staining and flow cytometry, the apoptosis by acridine orange/ ethidium bromide staining, and the cell senescence by beta-galactosidase staining; the levels of tumor necrosis factor alpha (TNF-alpha), interleukin 1 (IL-1), platelet-derived growth factor (PDGF), and insulin-like growth factor 1 (IGF-1) in serum were detected by a double-antibody sandwich ELISA kit.
hUCMSCs were long spindle/polygon in 3 groups. The cell fusion of group C was obviously faster than that in group A and group B. The cell proliferation curves showed that the velocity and number of cell proliferation in group C were significantly higher than those in group A and group B at 2-6 days after culture (P < 0.05). The rates of proliferation period (S) of hUCMSCs were 9.21% +/- 1.02%, 11.79% +/- 1.87%, and 20.54% +/- 2.03%, respectively in groups A, B, and C at 6 days, and group C was significantly higher than that of group A and group B (P < 0.05). The hUCMSCs showed normal morphology and structure in 3 groups, and no apoptosis cells was observed. The positive cells percentage of group C (2.6% +/- 0.1%) was significantly lower than that of group A (4.8% +/- 0.2%) and group B (3.8% +/- 0.4%) (P < 0.05). The levels of TNF-alpha, IL-1, PDGF, and IGF-1 in group C were significantly higher than those in group B (P < 0.05).
The higher levels of cytokines in serum from the severe burn patients can significantly stimulate hUCMSCs proliferation, prevent cells apoptosis, and reduce cells senescence. Therefore, it is feasible to use hUCMSCs transplantation for treating severe burn patients.
探讨重度烧伤患者血清对人脐带间充质干细胞（hUCMSCs）生物学特性的体外影响，以探索hUCMSCs移植治疗重度烧伤的可行性。
将第3代hUCMSCs随机分为3组：10%胎牛血清组（A组）、10%正常血清组（B组）和10%烧伤血清组（C组）。培养24小时、72小时和6天后，用倒置显微镜观察细胞形态和密度；用MTT法评估细胞增殖；培养6天后，用碘化丙啶染色和流式细胞术检测细胞周期，用吖啶橙/溴化乙锭染色检测细胞凋亡，用β-半乳糖苷酶染色检测细胞衰老；用双抗体夹心ELISA试剂盒检测血清中肿瘤坏死因子α（TNF-α）、白细胞介素1（IL-1）、血小板衍生生长因子（PDGF）和胰岛素样生长因子1（IGF-1）的水平。
3组hUCMSCs均呈长梭形/多边形。C组细胞融合明显快于A组和B组。细胞增殖曲线显示，培养2-6天后，C组细胞增殖速度和数量均显著高于A组和B组（P<0.05）。培养6天时，A组、B组和C组hUCMSCs的增殖期（S）比例分别为9.21%±1.02%、11.79%±1.87%和20.54%±2.03%，C组显著高于A组和B组（P<0.05）。3组hUCMSCs形态和结构正常，未观察到凋亡细胞。C组阳性细胞百分比（2.6%±0.1%）显著低于A组（4.8%±0.2%）和B组（3.8%±0.4%）（P<
