慢病毒载体感染人子宫内膜组织块的研究

目的在优化人子宫内膜组织块体外培养条件基础上,建立慢病毒载体感染人子宫内膜组织块的方法。方法将人子宫内膜标本剪成1 mm×1 mm×2 mm大小,移入含激素(雌二醇和醋酸甲羟孕酮)或不含激素的培养液中培养7 d,每天取3块组织块,进行大体和HE染色形态学观察。在优化的组织块体外培养条件下,组织培养液分别加入不同病毒量的慢病毒和具有促进病毒感染的感染增强液ENi.S和Polybrene,在荧光显微镜下观察荧光表达情况,比较阳性细胞率。结果人子宫内膜组织块在含激素的培养液中培养5 d后,色泽鲜红,结构完整,培养效果明显优于不含激素的培养液。3.5×10~7TU/ml病毒数量及在培养基中加入细胞增强液时,慢病毒感染阳性细胞数明显增加,阳性率达50%。结论慢病毒载体在体外可成功转染人子宫内膜组织块。

Transduction of cultured human endometrial tissues with lentivirus vector

Objective： To explore the effective and stable transduction of cultured human endometrial tissues with lentivirus vector. Methods： Human endometrial biopsies were cut into small pieces （1 mm × 1 mm × 2 mm） and maintained in culture medium with or without 1713-estradiol （Ez） plus medroxyprogesterone （MPA） for 7 days. 3 pieces of tissues were taken for gross observation and histological examination with HE. staining every day. The lentiviruses tagged with GFP were added into the medium at three different gradients with enhanced infection solution ENi. S and polybrene. The infection efficiency was identified by the expression of GFP observed with fluorescence microscope. Results： The tissues cultured with E2 plus MPA had better histological structures than those without supplementation of Ez and MPA. With addition of enhanced infection solution into the medium, the optimal lentivirus virus number for infecting human endometrial tissues was 3.5 × 10^7 TU/ml, which could achieve an infection efficiency of 50%. Conclusions： The model of infecting cultured human endometrial tissues with lentivirus vector was successfully established.