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无血清培养基中豚鼠植入前胚胎的发育

Development of preimplantation guinea-pig embryos in serum-free media.

作者信息

Suzuki O, Ogura A, Asano T, Noguchi Y, Yamamoto Y, Oike M

机构信息

Department of Veterinary Science, National Institute of Health, Tokyo, Japan.

出版信息

Reprod Fertil Dev. 1993;5(4):425-32. doi: 10.1071/rd9930425.

Abstract

This study was undertaken to determine how far preimplantation guinea-pig embryos can develop normally in vitro with serum-free media generally used for mouse (Whitten's medium, WM) and rabbit (Kane's medium, KM) embryos. Preimplantation guinea-pig embryos were recovered from naturally mated females at each day of pregnancy between Day 1 and Day 6, and were cultured in vitro with either WM or KM at 37 degrees C in 5% CO2 in air. One-cell (Day-1) and 2-cell (early Day 2) embryos cultured in WM developed to the 4-cell and morulae stages respectively, whereas the corresponding embryos cultured in KM arrested development at the 2-cell and 8-cell stages respectively. In contrast, 4-cell (late Day 2 and Day 3) embryos proceeded to the blastocyst stage in KM, but not in WM. Eight-cell embryos (Day 4) and morulae (Day 5) developed to blastocysts in either medium. However, hatching of the blastocysts and trophoblast outgrowth on the culture dish were supported only by KM even when collected at the blastocyst stage. The present study suggests that, in guinea-pig embryos, a distinct change in nutritional requirements occurs around the time of the second cell division and that, unlike in the mouse, trophoblast outgrowth in the guinea-pig requires no supplementation of serum or extracellular matrices.

摘要

本研究旨在确定植入前豚鼠胚胎在通常用于小鼠(惠顿培养基,WM)和兔子(凯恩培养基,KM)胚胎的无血清培养基中体外正常发育的程度。在妊娠第1天至第6天的每一天,从自然交配的雌性豚鼠中回收植入前胚胎,并在37℃、含5%二氧化碳的空气中用WM或KM进行体外培养。在WM中培养的单细胞(第1天)和2细胞(第2天早期)胚胎分别发育到4细胞和桑椹胚阶段,而在KM中培养的相应胚胎分别在2细胞和8细胞阶段停止发育。相反,4细胞(第2天晚期和第3天)胚胎在KM中发育到囊胚阶段,但在WM中则不能。8细胞胚胎(第4天)和桑椹胚(第5天)在两种培养基中均发育到囊胚阶段。然而,即使在囊胚阶段收集,囊胚的孵化和滋养层在培养皿上的生长也仅由KM支持。本研究表明,在豚鼠胚胎中,营养需求在第二次细胞分裂前后发生明显变化,并且与小鼠不同,豚鼠的滋养层生长不需要补充血清或细胞外基质。

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