A. I. Mazurkiewicz, V. Kovpak, O. S. Kovpak
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摘要

骨髓是唯一的成体组织,通常由未成熟的未分化和低分化的细胞组成,这些细胞被称为干细胞,它们在结构上与胚胎干细胞相似。但文献数据分析并没有给出大鼠骨髓细胞体外培养过程中表型和形态变化的明确答案,需要进一步研究。观察第1 ~ 4代大鼠骨髓细胞体外培养过程中表型和形态学的变化。我们在这些研究中使用了大鼠骨髓细胞从第一到第四代。利用倒置显微镜Axiovert 40对大鼠骨髓细胞培养过程中的形态学变化进行了显微镜分析和评价。通过检测CD标记物(CD10、CD38、CD34、CD45、CD48、CD54、CD56、CD66e、CD96、CD227、CD326、泛角蛋白)来控制表型变化。采用半定量方法(H-Score)进行评价。对大鼠骨髓细胞原代培养的研究表明,其形态不均匀,注意到少量细胞呈多角形,被成纤维细胞包围。在培养过程中,以牺牲成纤维细胞样细胞为代价,细胞培养变得更加均匀。由于发生了从零代异质文化到四代最同质文化的过渡过程。对大鼠骨髓培养的细胞进行免疫表型分析,发现泛角蛋白高水平表达;中等水平- CD34、CD48、CD66e、CD95;低水平- CD38、CD45、CD56、CD227、CD326;缺乏表达- CD10, CD54。各传代表面标记物CD48、CD66e、CD95表达变化显著增加;CD38, SD45, SD326,泛角蛋白明显减少。标记物CD34、cd56、cd227在第1 ~第4代呈一级表达。研究期间未发现CD10、CD54标记物的表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ФЕНОТИПОВІ ТА МОРФОЛОГІЧНІ ЗМІНИ КУЛЬТУРИ КЛІТИН КІСТКОВОГО МОЗКУ ЩУРІВ В ПРОЦЕСІ ЇХ КУЛЬТИВУВАННЯ
Bone marrow is the only adult tissue which normally consists of immature undifferentiated and low differentiated cells which called stem cells and they are similar in structure to embryonic stem cells. But literature data analysis doesn't give an unambiguous answer regarding phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation which necessitated further research. Investigate phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation from first to fourth passage. We were used in these research bone marrow cells of rats from the first to the fourth passages. Microscopic analysis and evaluation morphological changes of bone marrow cells culture of rats during cultivation were carried out using inverted microscope Axiovert 40. Control of changes phenotype was performed by detecting CD markers (CD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, pan–keratin). The evaluation was performed by the semi– quantitative method (H–Score). The research of primary culture of rat bone marrow cells showed that it morphologically heterogeneous, noted the small number of cells polygonal shape, surrounded by the fibroblast cells. During the cultivation cell culture becomes more homogenous at the expense of fibroblast–like cells. As a result of occurred the transition process from heterogeneous culture in zero passage to the most homogeneous culture in 4 passage. Immunophenotyping population of cell culture derived from rat bone marrow, revealed a high level of expression of pan–keratin; moderate level – CD34, CD48, CD66e, CD95; low level – CD38, CD45, CD56, CD227, CD326; lack of expression – CD10, CD54. Change of the expression of surface markers varies in each passage CD48, CD66e, CD95 increased significantly; CD38, SD45, SD326, pan–keratin reduced significantly. The markers CD34, CD 56, CD 227 were expressed on the one level from the first to the fourth passage. The expression of the CD10, CD54 markers during the study period was not identified.
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