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.