人正常宫颈上皮细胞的体外分离及细胞活性比较

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目的:在传统上皮细胞消化分离方法基础上加以比较,探求一种最佳宫颈上皮细胞的体外分离方法。方法:取10例新鲜的正常宫颈组织,将同一组织分成大小相同的3部分,采用3种不同的消化方法(胰蛋白酶消化法、Ⅰ型胶原酶消化法、Ⅱ型中性蛋白酶+胰蛋白酶联合消化法)获取正常宫颈上皮角质细胞悬液,采用罗丹明B(SRB)法测定其细胞活性,同时比较其贴壁率。结果:胰蛋白酶组、Ⅰ型胶原酶组、Ⅱ型中性蛋白酶+胰蛋白酶联合消化组的细胞活性分别为0.9335、0.9544、0.9438,两两比较差异显著(P<0.01);Ⅰ型胶原酶消化组的细胞贴壁率显著高于其他两组(P<0.01)。结论:Ⅰ型胶原酶消化法是一种最理想的分离宫颈上皮细胞的方法。 OBJECTIVE: To compare the traditional method of epithelial cell digestion and separation and to explore a method of in vitro isolation of the best cervical epithelial cells. Methods: Ten fresh normal cervical tissues were divided into three groups of the same size. Three different digestion methods (trypsin digestion, type Ⅰ collagenase digestion, type Ⅱ neutral protease + trypsin union Digestion method) to obtain normal cervical epithelial keratinocyte suspension, rhodamine B (SRB) method was used to determine the cell viability, while comparing the rate of attachment. Results: The cell viability of trypsin group, type Ⅰ collagenase group, type Ⅱ neutral proteinase + trypsin combined with digestion group were 0.9335, 0.9544 and 0.9438, respectively, with significant differences (P <0.01); type Ⅰ collagenase digestion Cell attachment rate was significantly higher than the other two groups (P <0.01). Conclusion: Type Ⅰ collagenase digestion is one of the most ideal method to isolate cervical epithelial cells.
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