Establishment of trophoblast cell line derived from buffalo (Bubalus bubalis) parthenogenetic embryo.

IF 1.5 4区 生物学 Q4 CELL BIOLOGY
Zygote Pub Date : 2024-08-01 Epub Date: 2024-10-14 DOI:10.1017/S0967199424000339
Sushil K Mohapatra, Anjit Sandhu, Prabhat Palta, Manoj K Singh, Suresh K Singla, Manmohan S Chauhan
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引用次数: 0

Abstract

We have established trophoblast cell lines, from parthenogenesis-derived buffalo blastocysts. The buffalo trophoblast cells were cultured continuously over 200 days and 21 passages. These cells were observed by phase-contrast microscopy for their morphology and characterized by reverse transcriptase polymerase chain reaction and immunofluorescence against trophoblast-specific markers and cytoskeletal proteins. Trophoblast cells showed positive staining for CDX2, a marker of these cells at both blastocyst and cell line levels. Epithelial morphology of these cells was revealed by positive staining against cytokeratins and tubulin but not against vimentin and dolichos biflorus agglutinin. Gene expression profiles of many important placenta-specific genes were studied in the primary trophectoderm outgrowths, which were collected on days 0, 5, 9, 12 and 15 of culture and trophoblast cell line at passages 12-15. Therefore, the trophoblast cell line derived can potentially be used for in vitro studies on buffalo embryonic development.

从水牛(Bubalus bubalis)孤雌胚胎中建立滋养层细胞系。
我们从孤雌生殖衍生的水牛囊胚中建立了滋养层细胞系。水牛滋养层细胞连续培养了 200 天和 21 个传代。这些细胞通过相位对比显微镜观察其形态,并通过逆转录酶聚合酶链反应和针对滋养层特异性标记物和细胞骨架蛋白的免疫荧光鉴定其特征。滋养层细胞对 CDX2 呈阳性染色,CDX2 是这些细胞在胚泡和细胞系水平上的标记物。细胞角蛋白和微管蛋白的阳性染色显示了这些细胞的上皮形态,但波形蛋白和双叶托叶凝集素的染色则没有显示。研究了在培养第 0、5、9、12 和 15 天收集的原始滋养层外胚层和第 12-15 代滋养层细胞系中许多重要的胎盘特异基因的基因表达谱。因此,所获得的滋养层细胞系可用于水牛胚胎发育的体外研究。
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来源期刊
Zygote
Zygote 生物-发育生物学
CiteScore
1.70
自引率
5.90%
发文量
117
审稿时长
6-12 weeks
期刊介绍: An international journal dedicated to the rapid publication of original research in early embryology, Zygote covers interdisciplinary studies on gametogenesis through fertilization to gastrulation in animals and humans. The scope has been expanded to include clinical papers, molecular and developmental genetics. The editors will favour work describing fundamental processes in the cellular and molecular mechanisms of animal development, and, in particular, the identification of unifying principles in biology. Nonetheless, new technologies, review articles, debates and letters will become a prominent feature.
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