人少突胶质前体细胞传代过程中形态学的变化

Morphological changes in human oligodendrocyte progenitor cells during passage

背景:少突胶质前体细胞是治疗脑白质损伤的种子细胞,建立高效稳定的体外培养方法是临床应用的重要前提。目的:探讨少突胶质前体细胞培养过程中的形态变化。方法:采用4批人少突胶质前体细胞从第2代体外传代培养至第7代,每次传代前在光学显微镜下拍200倍视野照片5张,根据细胞形态将少突胶质前体细胞分为双极类细胞(少突胶质前体细胞)、多极类细胞(晚期少突胶质前体细胞)和极上分叉类细胞(未成熟少突胶质细胞)3个类型,分别计算各类型细胞占总细胞数的比例,从而比较各个代数之间细胞形态有无差异。结果与结论:少突胶质前体细胞从第2代传代至第7代过程中,包含双极类细胞、多极类细胞和极上分叉类细胞3种类型,其中以双极类细胞和多极类细胞为主,剩余可见少量极上分叉类细胞。各代细胞中的双极类细胞比例、多极类细胞比例及极上分叉类细胞比例差异无显著性意义(P>0.05)。结果表明,采用细胞形态分类计数方法初步评估得出少突胶质前体细胞在培养过程中形态无变化。

BACKGROUND:Oligodendrocyte precursor cells are seed cells for the treatment of white matter injury.The establishment of an efficient and stable in vitro culture method is an important prerequisite for clinical transformation.OBJECTIVE:To investigate the morphological changes of oligodendrocyte precursor cells during passage.METHODS:Four batches of human oligodendrocyte progenitor cells were subcultured from the second generation(P2)to the seventh generation(P7)in vitro.Five pictures of 200-fold field were taken under an optical microscope before each passage.According to the cell morphology,oligodendrocyte precursor cells were divided into three types:bipolar cells(oligodendrocyte precursor cells),multipolar cells(late oligodendrocyte precursor cells)and supra-polar bifurcated cells(immature oligodendrocyte cells).The proportion of oligodendrocyte progenitor cells to the total number of cells was calculated,so as to compare the difference of cell morphology among different generations.RESULTS AND CONCLUSION:In the process of passage from P2 to P7,oligodendrocyte progenitor cells included three types:bipolar cells,multipolar cells and supra-polar bifurcated cells.Among them,bipolar cells and multipolar cells were the main part,and a small number of supra-polar bifurcated cells could be seen in the rest.There were no significant differences in the proportion of bipolar cells,multipolar cells and supra-polar bifurcated cells among P2-P7(P>0.05).The cell morphology classification and counting method can be used to preliminarily evaluate that oligodendrocyte progenitor cells have no change in morphology during culture.