应用流式细胞术测定CD24表达鉴定人髓核细胞

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。

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

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.