汪兆艳,王 倩,杨印祥,屈素清,杨 辉,叶 豆,门倩倩,刘 畅,栾 佐.冷藏保存时间对人少突胶质前体细胞的影响[J].转化医学杂志,2022,11(5):269-273
冷藏保存时间对人少突胶质前体细胞的影响
Impact of cryopreservation time on biological properties of human oligodendrocyte precursor cells
  
DOI:
中文关键词:  少突胶质前体细胞  保存介质  细胞分化  细胞活率
英文关键词:Oligodendrocyte precursor cell  Storage medium  Differentiation  Proliferation  Viability
基金项目:国家重点研发计划(2017YFA0104200)
作者单位
汪兆艳 解放军总医院第六医学中心儿科 
王 倩 解放军总医院第六医学中心儿科 
杨印祥 解放军总医院第六医学中心儿科 
屈素清 解放军总医院第六医学中心儿科 
杨 辉 解放军总医院第六医学中心儿科 
叶 豆 解放军总医院第六医学中心儿科 
门倩倩 解放军总医院第六医学中心儿科 
刘 畅 解放军总医院第六医学中心儿科 
栾 佐 解放军总医院第六医学中心儿科 
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中文摘要:
      目的 评价冷藏保存时间对人少突胶质前体细胞生物学特性的影响。方法 取生长状态良好的第3代人少突胶质前体细胞,按照4×107 mL-1细胞浓度重悬于含有10 g/L 人血清白蛋白的50 g/L葡萄糖保存介质中,置于2~8 ℃冷链运输箱中保存24 h、48 h、72 h,与新鲜制备的少突胶质前体细胞(即冷藏保存0 h细胞)比较,评价冷藏保存时间对细胞活率、增殖能力、迁移能力、免疫表型及分化能力的影响。将冷藏保存24 h的少突胶质前体细胞移植到脑白质损伤大鼠侧脑室内,观察移植细胞在体内成髓鞘情况。结果 与冷藏保存0 h细胞比较,冷藏保存24 h、48 h细胞活率没有显著差异,冷藏保存72 h细胞活率明显下降(P<0.05);通过CCK8法检测各组细胞增殖能力,结果显示冷藏保存24 h和48 h细胞增殖能力与冷藏保存0 h的细胞比较无显著差异,冷藏保存72 h细胞增殖能力显著下降(P<0.05);细胞体外迁移和分化结果显示,与冷藏保存0 h细胞比较,冷藏保存24 h的细胞迁移和分化能力无显著差异,冷藏保存48 h和72 h,细胞迁移和分化能力显著下降。体内实验证实,冷藏保存24 h的少突胶质前体细胞在体内可以分化为产髓鞘碱性蛋白的少突胶质细胞。结论 人少突胶质前体细胞在含有10 g/L人血白蛋白的50 g/L葡萄糖保存介质中冷藏保存24 h,细胞活率、迁移和分化能力较好,因此,建议少突胶质前体细胞制剂冷藏保存24 h内移植,以确保最佳治疗效果。
英文摘要:
      Objective To evaluate the impact of cryopreservation time on biological properties of human oligodendrocyte precursor cells (OPCs).Methods Passage 3 human OPCs in normal growth were re-suspended in 50g/L glucose medium containing 10g/L human serum albumin (HSA) at a concentration of 4X107mL-1 and stored in the 2~8℃cold-chain transport box for 24, 48 and 72 h, and were compared to freshly prepared oligodendrocyte precursor cells(OPCs stored for 0 h. Then, OPCs stored for 24 h were transplanted to the brain lateral ventricle of rats with white matter injury (WMI) to observe their myelination in vivo. Results Compared with the cells stored for 0 h, there was no significant difference in the viability of cells stored for 24 h and 48 h, but cell viability of OPCs stored for 72 h decreased significantly (P<0.05). The result of CCK8 assay showed that there was no significant difference in cell proliferation between OPCs stored for 24 and 48 h and those stored for OPCs stored for 0 h, but cell proliferation of OPCs stored for 72 h was decreased significantly (P<0.05). The results of cell migration and differentiation in vitro showed that compared with 0 h cells, there was no significant difference in cell migration and differentiation ability at 24 h, but significantly decreased at 48 h and 72 h.In vitro migration and differentiation experiments showed that the migration and differentiation abilities of OPCs decreased significantly with the increase of the length of cryopreservation time. The in vivo experiment demonstrated that OPCs stored for 24 h could differentiate to myelin basic protein (MBP)-positive oligodendrocytes.Conclusion Human oligodendrocyte precursor cells are refrigerated in 50g/L glucose storage medium containing 10g/L human albumin for 24 h, and the cell viability, migration and differentiation ability are the best. Therefore, it is recommended that oligodendrocyte precursor cells The preparation should be refrigerated and transplanted within 24 hours to ensure the best therapeutic effect.
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