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培养的兔下颌髁突软骨细胞的表型变化。

Phenotypic changes of rabbit mandibular condylar cartilage cells in culture.

作者信息

Engel F E, Khare A G, Boyan B D

机构信息

Department of Orthodontics, University of Texas Health Science Center, San Antonio 78284.

出版信息

J Dent Res. 1990 Nov;69(11):1753-8. doi: 10.1177/00220345900690110801.

Abstract

The present study describes the behavior of mandibular condylar cartilage (MCC) cells as a function of time in primary culture, since it is not yet clear whether these cells maintain their phenotype in culture. MCC cells from New Zealand white rabbits were seeded at high density and cultured in DMEM containing 50 micrograms/mL ascorbic acid and 10% fetal bovine serum. These cells appeared as a heterogeneous population and changed their shape, size, and refractivity as cultures aged. Cartilage-like cells, which always dominated the culture, were infiltrated with a minority of fibroblast-like cells. Cell number increased progressively, and cultures reached confluence at nine days. Antibody activity for cartilage-specific glycosaminoglycan was determined by ELISA assay. This reaction reached a maximum at six days and decreased thereafter. Cultures stained with Alcian blue (pH 1.0) supported these results. Cytoplasmic mRNA analysis indicated that the transcription of type II collagen gene was present at all time points. Type I collagen and alkaline phosphatase mRNA levels showed progressive increases from 12 h to nine days, with significantly higher values in cells cultured for six, nine, and 12 days than in cells collected from earlier time points. These results suggest that in our present culture system, MCC cells undergo phenotypic changes that resemble their maturation processes in vivo.

摘要

由于目前尚不清楚下颌髁突软骨(MCC)细胞在培养过程中是否能保持其表型,本研究描述了原代培养中MCC细胞随时间变化的行为。将来自新西兰白兔的MCC细胞高密度接种,并在含有50微克/毫升抗坏血酸和10%胎牛血清的DMEM中培养。随着培养时间的延长,这些细胞呈现出异质性群体,其形状、大小和折光率都发生了变化。始终占主导地位的软骨样细胞中混入了少数成纤维细胞样细胞。细胞数量逐渐增加,培养物在第9天达到汇合。通过ELISA测定法测定软骨特异性糖胺聚糖的抗体活性。该反应在第6天达到最大值,此后下降。用阿尔辛蓝(pH 1.0)染色的培养物支持了这些结果。细胞质mRNA分析表明,II型胶原基因的转录在所有时间点均存在。I型胶原和碱性磷酸酶mRNA水平从12小时到9天呈逐渐增加趋势,培养6天、9天和12天的细胞中的值明显高于早期时间点收集的细胞。这些结果表明,在我们目前的培养系统中,MCC细胞经历了类似于其体内成熟过程的表型变化。

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