【摘要】 目的观察低温冻存1月后,不同年龄大鼠髁突软骨细胞的生物学特性。方法体外培养出生后1天、7天、14天、28天共4组SD大鼠髁突软骨细胞。对冻存复苏后的细胞进行形态学观察;甲苯胺蓝染色、Ⅱ型胶原免疫组化染色检测其对糖胺聚糖(GAG)、Ⅱ型胶原的合成能力;台盼蓝拒染试验测定细胞成活率;MTT法观察细胞增殖能力;实时定量-聚合酶链式反应(real time-PCR)检测冻存后细胞增殖核抗原(PCNA)基因表达情况。结果低温冻存后,各年龄组大鼠髁突软骨细胞仍能保持软骨细胞特有的形态及特性;细胞成活率均大于90%;生长曲线近似"S"型;冻存后各年龄组细胞pcna基因表达较冻存前无显著统计学差异(P>0.05)。结论低温冻存1月后的大鼠髁突软骨细胞成活率高,并保持了其生物学特性及增殖活性。更多还原

【Abstract】 Objective To study the biological characteristics of rat condylar chondrocytes after 1 month frozen-thawing.Methods The 1,7,14 and 28 day-old SD rat condylar chondrocytes were cultured in vitro.The morphology of frozen-thawed chondrocytes was observed,and their ability to synthesizing glycosaminoglycans(GAG) and type Ⅱ collagen was determined by toluidine blue stain and type Ⅱ collagen immunohistochemistry reaction.Viability and the proliferation activity were detected by trypan blue exclusion method,MTT assay.Otherwise,real time-PCR was performed by using proliferation cell nuclear antigen(PCNA) specific primers to detect its gene expresion.Results The frozen-thawed condylar chondrocytes could still keep the morhoplogy and biological characteristics of cartilage chondrocytes specially;Cell viability was over 90%.The growth curve of condylar chondrocytes resembled "S" in shape.Proliferation cell nuclear antigen(PCNA) gene expression changed little between the before-and after-cryopresevation condylar chondroctyes(P>0.05).Conclusion Rat condylar chondrocytes after 1 month frozen-thawing still maintain their biological characteristics,proliferation activity and keep high viability.更多还原