人少突胶质前体细胞在不同细胞培养器皿中的比例和形态特征

Proportion and morphological characteristics of human oligodendrocyte precursor cells in different cell culture vessels

背景:少突胶质前体细胞移植是治疗早产儿脑白质损伤的关键措施之一,目前国内外缺乏人胎脑来源神经干细胞诱导的少突胶质前体细胞在不同器皿中培养比较的研究。目的:观察不同细胞培养器皿(6孔板、24孔板和T25培养瓶)对人胎脑来源神经干细胞系诱导培养的人少突胶质前体细胞和前-少突胶质细胞形态的影响。方法:采用6孔板、24孔板和T25培养瓶作为培养器皿培养人少突胶质前体细胞和前-少突胶质细胞,免疫荧光染色和流式细胞术对人少突胶质前体细胞的特性进行鉴定,普通光学显微镜观察细胞形态,依据细胞形态进行计数并统计分析。结果与结论:①少突胶质前体细胞胞体呈圆形,双极突起呈串珠样结构;前-少突胶质细胞突起多于2极但不分叉;②与T25培养瓶、24孔板相比,6孔板中双极串珠样的少突胶质前体细胞比例最高,差异有显著性意义(P <0.05),其次依次为T25培养瓶、24孔板;与T25培养瓶、6孔板相比,24孔板中前-少突胶质细胞的比例最高,差异有显著性意义(P <0.01),其次依次为T25培养瓶、6孔板;③6孔板中培养的细胞渣滓少,形态、活力和长势比其他培养皿更佳;④从形态学角度分析,6孔板更适宜于少突胶质前体细胞的生长,24孔板更适宜于前-少突胶质细胞的生长。

BACKGROUND:Oligodendrocyte precursor cell transplantation is one of the keys to the treatment of white matter damage in premature infants.At present,there is a lack of research on the comparison of oligodendrocyte precursor cells induced by neural stem cells derived from human fetal brain cultured in vitro in different vessels worldwide.OBJECTIVE:To observe the morphology of human oligodendrocyte progenitor cells and pre-oligodendrocytes in different cell culture vessels(6-well plates,24-well plates and T25 flasks).METHODS:The 6-well plates,24-well plates and T25 flasks were used as culture vessels to culture human oligodendrocyte progenitor cells and preoligodendrocytes.The characteristics of human oligodendrocyte progenitor cells were identified by immunofluorescence staining and flow cytometry.The morphology of cells was observed by an ordinary light microscope.Cell counts were performed according to cell morphology and statistical analysis was performed.RESULTS AND CONCLUSION:(1)The oligodendrocyte progenitor cell body was round and the bipolar protrusions were bead-like;the pre-oligodendrocyte protrusions were more than two poles,and did not bifurcate.(2)The ratios of oligodendrocyte progenitor cell morphology in the oligodontia lines were significantly higher in the 6-well plates than those in the 24-well plates and T25 flasks(P<0.05),followed by T25 flasks and 24-well plates.Morphological ratios of pre-oligodendrocytes were significantly higher in the 24-well plates compared to the 6-well plates and T25 flasks(P<0.01),followed by T25 flasks and 6-well plates.(3)The cells cultured in the 6-well plate had fewer dregs,and the morphology,vigor and growth were better than those of the other culture vessels.(4)According to morphological view,6-well plates are more suitable for oligodendrocyte progenitor cell growth,and 24-well plates are more suitable for preoligodendrocytes growth.