绵羊绒毛膜滋养层细胞与子宫内膜腔上皮细胞共培养体系的建立

doi:10.11843/j.issn.0366-6964.2019.03.011

Abstract

Abstract:

The study aims to explore the co-culture conditions of sheep trophoblast cells (STCs) and the endometrial luminal epithelial cells (ELECs), and provides experimental basis for chorionic trophoblastic coenocyte formation. Harvesting healthy sheep uterus and placental tissue from 45 to 60 days of gestation at the slaughter house, and the enzyme digestion method has been used to isolate and culture the STCs and ELECs in vitro, and they were identified by cellular immunofluorescence. Simultaneously, the optimal co-culture conditions for STCs and ELECs were determined by Giemsa staining. Then, the empty plasmid pEGFP-C1 was transfected into ELECs for 48 h, and then co-cultured with STCs. The morphology of chorionic trophoblastic coenocyte was observed by confocal laser scanning microscopy. The results showed that STCs CK-7 and ELECs CK-18 were positive.When the ratio of STCs to ELECs cells was 2:1, the number of multinucleated cells reaches the maximum when the co-culture time was 48 h, and it was significantly different from the control group (P<0.05). The results of immunofluorescence showed that CK-7 expressed red fluorescence in STCs, and there were pEGFP-C1 labeled green fluorescence in the cells, and the nucleus was blue-stained, and the 3 fluorescent co-expressions showed white light. This study established the conditions for co-culture of STCs and ELECs, and showed that ELECs were involved in the formation of polynuclear cells in sheep trophoblasts.

CLC Number:

S826.2

ZHANG Jie, CHEN Dayong, ZHAO Juan, WANG Xiaojuan, YANG Hui, LIU Shuying. The Establishment of Co-culture System of Sheep Trophoblast Cells and Endometrial Luminal Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 562-569.

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The Establishment of Co-culture System of Sheep Trophoblast Cells and Endometrial Luminal Epithelial Cells

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