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猪早期胚胎与输卵管上皮细胞共培养的体外发育

Early embryonic development in vitro by coculture with oviductal epithelial cells in pigs.

作者信息

White K L, Hehnke K, Rickords L F, Southern L L, Thompson D L, Wood T C

机构信息

Department of Animal Science, LSU Agricultural Center, Louisiana State University, Baton Rouge 70803.

出版信息

Biol Reprod. 1989 Sep;41(3):425-30. doi: 10.1095/biolreprod41.3.425.

Abstract

This experiment was designed to evaluate the ability of three different somatic cell cultures to promote development of early cleavage stage pig embryos. A total of 245 2-cell, 4-cell, 8-cell, and 16-cell pig embryos were cocultured for 5 days with porcine oviductal epithelial cells (POEC), porcine fetal fibroblast monolayer (PEF), a combined POEC and PEF coculture system (PEF-POEC), or Dulbecco's Modified Eagle Medium alone (DMEM). Embryos were collected at slaughter from the reproductive tracts of superovulated prepubertal gilts. Embryos were recovered, evaluated, and randomly placed in one of the four treatment groups. POEC were recovered from oviductal flushes, washed, and placed in 24-well plates. PEF were obtained from 30-day to 60-day fetuses and established in culture. Finally, PEF-POEC consisted of a confluent monolayer of PEF in the bottom of 24-well plates also containing a Costar semipermeable membrane chamber with POEC in it. Embryos were evaluated every 24 h to determine stage of development. More (p less than 0.05) embryos developed to blastocysts in POEC (70% and 54%, respectively) and PEF-POEC (67% and 61%, respectively), than in either DMEM (16% and 2%, respectively) or PEF (27% and 23%, respectively). However, development of embryos did not differ (p less than 0.05) for POEC and PEF-POEC. These data indicate the presence of a primary culture of POEC promotes in vitro development of early cleavage stage pig embryos.

摘要

本实验旨在评估三种不同体细胞培养物促进猪早期卵裂期胚胎发育的能力。将总共245个2细胞、4细胞、8细胞和16细胞的猪胚胎与猪输卵管上皮细胞(POEC)、猪胎儿成纤维细胞单层(PEF)、POEC和PEF联合共培养系统(PEF-POEC)或单独的杜氏改良 Eagle 培养基(DMEM)共培养5天。胚胎从超排未成熟小母猪的生殖道中屠宰采集。胚胎被回收、评估,并随机置于四个处理组之一。POEC从输卵管冲洗液中回收、洗涤后置于24孔板中。PEF取自30至60天龄的胎儿并进行培养。最后,PEF-POEC由24孔板底部的PEF汇合单层组成,24孔板中还含有一个装有POEC的Costar半透膜小室。每24小时评估一次胚胎以确定发育阶段。与DMEM组(分别为16%和2%)或PEF组(分别为27%和23%)相比,POEC组(分别为70%和54%)和PEF-POEC组(分别为67%和61%)发育至囊胚的胚胎更多(p<0.05)。然而,POEC组和PEF-POEC组胚胎的发育情况并无差异(p<0.05)。这些数据表明,POEC原代培养物的存在可促进猪早期卵裂期胚胎的体外发育。

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