Effect of Induction Time on the Proliferation and Differentiation of Induced Schwann-Like Cells from Adipose-Derived Stem Cells.
Effect of Induction Time on the Proliferation and Differentiation of Induced Schwann-Like Cells from Adipose-Derived Stem Cells.
复制标题
诱导时间对脂肪干细胞诱导雪旺样细胞增殖和分化的影响。
DOI：
10.1007/s10571-020-00795-5
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发表时间：
2020
影响因子：
4
通讯作者：
He Bo
中科院分区：
文献类型：
--
作者：
Wong Chau Wei;Xu Yangbin;Liu Xiangxia;Xu Shuqia;Zhang Yi;Zhu Zhaowei;He Bo
为了比较不同诱导时间对脂肪干细胞（ADSC）诱导的许旺样细胞（iSC）增殖和分泌能力的影响，从健康成年雌性大鼠中分离ADSC。流式细胞术（FCM）检测ADSC阳性标志物CD 29、CD 44和CD 90以及阴性标志物CD 45。使用iSC诱导培养基培养ADSC。通过免疫荧光染色检测S-100、GFAP、MBP和P75以鉴定iSC分化。倒置显微镜下观察细胞形态学变化。MTS试验用于评估细胞增殖能力。应用Western blot分析caspase-3/切割的caspase-3和FCM评估细胞凋亡。建立PC 12与ADSCs或iSCs共培养体系，分析iSCs的生物学功能。在检测的蛋白质中，S-100、GFAP、MBP和P75在iSC中表达。第7天后，细胞增殖率明显低于诱导前，第19天，iSCs增殖率低于诱导前的50%（OD值= 0.016 ± 0.003 vs. 0.400 ± 0.004，p < 0.01）。从第19天开始，P21、P53、Apoj、S100、Gdnf和Mbp均一致地显示表达增加的趋势。NGF、MBP和BDNF的分泌在第19天较第7天增强。在共培养体系中，iSCs的诱导效果在第19天比第7天更明显，差异有统计学意义（55.40 ± 4.50 μm vs 37.15 ± 3.75 μm，p< 0.01）。总之，ADSC来源的iSC的增殖能力与诱导时间呈负相关，而SC标志蛋白的表达呈正相关。因此，iSC适合在诱导后19天使用。
To compare how different induction time takes effect on the proliferation and secretion ability of adipose-derived stem cell (ADSC)–induced Schwann-like cells (iSCs), ADSCs were isolated from healthy adult female rats. Flow cytometry (FCM) was performed to detect the ADSC-positive markers CD29, CD44, and CD90 and the negative marker CD45. iSC induction medium was used to culture the ADSCs. S-100, GFAP, MBP, and P75 were detected by immunofluorescence staining to identify iSC differentiation. Cell morphological changes were observed by an inverted microscope after induction. An MTS assay was used to evaluate the cell proliferation ability. Western blot analyses of caspase-3/cleaved caspase-3 and FCM were applied to assess cell apoptosis. Co-culture system of PC12 and ADSCs or iSCs was established to analyse the biological function of iSCs. Among the examined proteins, S-100, GFAP, MBP, and P75 were expressed in iSCs. After day 7, the cell proliferation rate was significantly lower than that before induction, and on day 19, the proliferation rate of iSCs was lower than 50% of the proliferation rate before induction (OD value = 0.016 ± 0.003 vs. 0.400 ± 0.004,p< 0.01). Starting from day 19, P21, P53, Apoj, S100, Gdnf, and Mbp all consistently showed a trend toward increased expression. Secretion of NGF, MBP, and BDNF was more enhanced at 19 days than that at 7 days. In co-culture system, the induction effect of iSCs was more pronounced at 19 days than that at 7 days, and the difference was statistically significant (55.40 ± 4.50 μm vs 37.15 ± 3.75 μm,p< 0.01). In conclusion, the proliferation ability of ADSC-derived iSCs was negatively correlated with the induction time, while the expression of SC marker proteins was positively correlated. Therefore, iSCs are suitable for use at 19 days after induction.