One-step versus two-step culture of mouse preimplantation embryos: is there a difference?
One-step versus two-step culture of mouse preimplantation embryos: is there a difference?
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DOI：
10.1093/humrep/dei228
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发表时间：
2005-12-01
影响因子：
6.1
通讯作者：
Lawitts, JA
中科院分区：
文献类型：
--
作者：
Biggers, JD;McGinnis, LK;Lawitts, JA
背景：比较了小鼠受精卵在一步法和两步法培养系统中的发育。方法：进行胚胎培养、囊胚细胞计数和胚胎移植。研究结果：在囊胚比例、孵化率、内细胞团（ICM）和滋养外胚层（TE）中的细胞数方面没有观察到显著差异，这些方案是：在补充葡萄糖和氨基酸的富钾单纯优化培养基（KSOMgAA）中的一步培养，在KSOMgAA/KSOMgAA中的两步培养，以及在G1.2/G2.2中的两步培养。在使用KSOMgAA的一步方案和使用G1.2/G2.2的两步方案中，从受精卵发育的胎仔中未观察到大体异常。这两组胎仔的体重无显著差异，未观察到发育异常。在KSOMgAA一步培养、KSOMgAA/KSOMgAA两步培养和DM 2/DM 1两步培养方案中，未观察到囊胚比例、孵化率、ICM和TE中细胞数的显著差异。EDTA在KSOMgAA中浓度为0.01 mmol/l时对发育的初始卵裂阶段无毒性。结论：两步培养方案足以支持植入前小鼠的体外发育，但它们不是必需的。
BACKGROUND: A comparison has been made of the development of mouse zygotes in either one-step or two-step culture systems. METHODS: Embryo culture, blastocyst cell counts and embryo transfer were done. RESULTS: No significant differences were observed in the proportions of blastocysts, rates of hatching, numbers of cells in the inner cell mass (ICM) and trophectoderm (TE) that developed in protocols: one-step culture in potassium-enriched simplex optimized medium supplemented with glucose and amino acids (KSOMgAA), two-step culture in KSOMgAA/KSOMgAA, and two-step culture in G1.2/G2.2. No gross abnormalities were observed in the fetuses that developed from zygotes in the one-step protocol using KSOMgAA and a two-step protocol using G1.2/G2.2. The body weights of these two groups of fetuses were not significantly different and no developmental abnormalities were observed. No significant differences were observed in the proportions of blastocysts, rates of hatching, numbers of cells in the ICM and TE that developed in protocols: one-step culture in KSOMgAA, two-step culture in KSOMgAA/KSOMgAA, and two-step culture in DM2/DM1. EDTA is not toxic to the initial cleavage stages of development at a concentration of 0.01 mmol/l in KSOMgAA. CONCLUSIONS: Two-step culture protocols are sufficient for the support of preimplantation mouse development in vitro but they are not necessary.