应用流式细胞术测定CD24表达鉴定人髓核细胞  

Identification of human nucleus pulposus cells by detecting CD24 expression using flow cytometry

作　　者：张丽[1] 马迅[1] 关晓明[1] 何李明[1] 赵胜[1] 宋文慧[1] 

机构地区：[1]山西医科大学第二医院骨科,山西省太原市030001

出　　处：《中国组织工程研究与临床康复》2011年第46期8583-8586,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:CD24是成熟髓核细胞表面表达的特异性分子,测定CD24的表达是否可以作为鉴定髓核细胞的一种方法?目的:验证流式细胞仪测定髓核细胞CD24表达鉴定髓核细胞的方法。方法:采用酶消化法分离培养人髓核细胞,取生长旺盛的第2代人髓核细胞爬片进行Ⅱ型胶原免疫组织化学染色,RT-PCR测定Ⅱ型胶原及SOX9的表达,流式细胞仪检测其CD24阳性表达率,分析来自同一批细胞的免疫组织化学染色、RT-PCR结果与CD24阳性率的相关性。结果与结论:体外分离培养的7例第2代髓核细胞爬片Ⅱ型胶原免疫组织化学染色呈阳性,RT-PCR结果阳性,流式细胞仪测定CD24其阳性率均为50%以上,髓核细胞分泌胞外基质Ⅱ型胶原与SOX9的表达与CD24阳性率结果有相关性。提示采用流式细胞仪测定CD24表达可以快速、简便地鉴定髓核细胞。BACKGROUND：CD24 is known as its specific expression on the surface of nucleus pulposus（NP） cells.Whether detecting CD24 expression can be used as a method to identify NP cells remains poorly understood.OBJECTIVE：To verify whether flow cytometry（FCM） detection of CD24 expression in NP cells is a fast and convenient method to identify NP cells.METHODS：Human NP cells were isolated and cultured from seven patients who underwent various spinal surgeries via enzyme digestion.Type Ⅱ collagen immunohistochemistry staining was proceeded to the well grown passage 2 NP cells.RT-PCR experiments had been done to measure the mRNA expression of type Ⅱ collagen and SOX9 from the same cell source.Also the positive rates of CD24 were detected by FCM.The correlation among these results from the same sample was analyzed.RESULTS AND CONCLUSION：Type Ⅱ collagen immunohistochemistry staining results of in vitro cultured passage 2 NP cells was positive.RT-PCR and FCM also acquired positive results.The positive rate of CD24 was above 50%.Extracellular matrix typeⅡcollagen and SOX9 expression of NP cells correlated to positive CD24 results.Detecting CD24 expression using FCM can identify NP cells fast and conveniently.

关 键 词：髓核细胞 流式细胞术 CD24 Ⅱ型胶原 SOX9 

分 类 号：R318[医药卫生—生物医学工程]