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目的:探索分离子宫内膜细胞以及纯化子宫内膜腺上皮细胞与间质细胞的最佳方法。方法:采用Ⅰ型胶原酶、胰酶、Ⅰ型胶原酶混合胰酶以及机械研磨等4种方法处理大鼠子宫,比较细胞数和活细胞率,从而获得分离子宫内膜组织的最佳方法。用获得的最佳方法分离人类子宫内膜组织,比较采用2次筛网过滤(筛网法)和低速离心结合筛网过滤(结合法)进一步纯化子宫内膜腺上皮细胞和间质细胞获得的细胞数和纯化效率,从而获得较好的纯化方法。结果:Ⅰ型胶原酶、胰酶、Ⅰ型胶原酶混合胰酶和机械研磨处理大鼠子宫组织所得的细胞数分别为6.43±3.55×105/ml、4.59±2.35×105/ml、4.25±1.06×105/ml、3.57±1.15×105/ml,差异有统计学意义(P<0.05);活细胞率分别为98.90±0.74%、96.63±1.84%、97.97±2.02%、97.20±4.41%,差异无统计学意义(P>0.05)。筛网法和结合法纯化分离所获得的腺上皮细胞数为0.43±0.21×105/ml、8.27±2.46×105/ml;间质细胞纯度为92.94±2.89%和99.19±0.24%,差异均有统计学意义(P<0.05)。结论:Ⅰ型胶原酶是分离子宫内膜细胞的最佳方法,而采用低速离心结合筛网过滤法是较理想的纯化分离后的子宫内膜细胞的方法。
Objective: To explore the best way to separate endometrial cells and purify endometrial glandular epithelial cells and interstitial cells. Methods: Uteri were treated with collagenase type Ⅰ, trypsin, type Ⅰ collagenase, trypsin and mechanical grinding. The best method of separating endometrial tissue was obtained by comparing the number of cells and viable cells. Human endometrial tissue was isolated using the best method obtained, and further purified by endometrial glandular epithelial cells and interstitial cells using secondary sieve filtration (sieve method) and low-speed centrifugation combined with sieve filtration (binding method) Cell number and purification efficiency, so as to obtain a better purification method. Results: The number of cells treated with type Ⅰ collagenase, trypsin, type Ⅰ collagenase and trypsin and mechanical grinding of uterine tissue were 6.43 ± 3.55 × 105 / ml, 4.59 ± 2.35 × 105 / ml and 4.25 ± 1.06 The difference was statistically significant (P <0.05). The viable cell rates were 98.90 ± 0.74%, 96.63 ± 1.84%, 97.97 ± 2.02% and 97.20 ± 4.41%, respectively No statistical significance (P> 0.05). The number of glandular epithelial cells obtained by sieving and purification was 0.43 ± 0.21 × 105 / ml and 8.27 ± 2.46 × 105 / ml, respectively. The purity of mesenchymal cells was 92.94 ± 2.89% and 99.19 ± 0.24%, respectively Statistical significance (P <0.05). Conclusion: Type Ⅰ collagenase is the best method for the separation of endometrial cells. The combination of low speed centrifugation and sieve filtration is an ideal method for the purification of isolated endometrial cells.