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商用胚胎培养液的组成。

Composition of commercial media used for human embryo culture.

机构信息

Department of Obstetrics and Gynecology, Mayo Clinic, Rochester, Minnesota; Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota.

National Foundation for Fertility Research, Lone Tree, Colorado.

出版信息

Fertil Steril. 2014 Sep;102(3):759-766.e9. doi: 10.1016/j.fertnstert.2014.05.043. Epub 2014 Jul 4.

Abstract

OBJECTIVE

To determine the composition of commercially available culture media and test whether differences in composition are biologically relevant in a murine model.

DESIGN

Experimental laboratory study.

SETTING

University-based laboratory.

ANIMAL(S): Cryopreserved hybrid mouse one-cell embryos were used in experiments.

INTERVENTION(S): Amino acid, organic acid, ions, and metal content were determined for two different lots of media from Cook, In Vitro Care, Origio, Sage, Vitrolife, Irvine CSC, and Global. To determine whether differences in the composition of these media are biologically relevant, mouse one-cell embryos were thawed and cultured for 120 hours in each culture media at 5% and 20% oxygen in the presence or absence of protein in an EmbryoScope time-lapse incubator.

MAIN OUTCOME MEASURE(S): The compositions of seven culture media were analyzed for concentrations of 39 individual amino acids, organic acids, ions, and elements. Blastocyst rates and cell cycle timings were calculated at 96 hours of culture, and the experiments were repeated in triplicate.

RESULT(S): Of the 39 analytes, concentrations of glucose, lactate, pyruvate, amino acids, phosphate, calcium, and magnesium were present in variable concentrations, likely reflecting differences in the interpretation of animal studies. Essential trace elements, such as copper and zinc, were not detected. Mouse embryos failed to develop in one culture medium and were differentially affected by oxygen in two other media.

CONCLUSION(S): Culture media composition varies widely, with differences in pyruvate, lactate, and amino acids especially notable. Blastocyst development was culture media dependent and showed an interaction with oxygen concentration and presence of protein.

摘要

目的

确定市售培养基于的组成,并检验其组成差异在小鼠模型中是否具有生物学相关性。

设计

实验性实验室研究。

设置

大学实验室。

动物

使用冷冻保存的杂交鼠单细胞胚胎进行实验。

干预

测定来自 Cook、In Vitro Care、Origio、Sage、Vitrolife、Irvine CSC 和 Global 的两种不同批次培养基中的氨基酸、有机酸、离子和金属含量。为了确定这些培养基组成差异是否具有生物学相关性,将小鼠单细胞胚胎解冻并在存在或不存在蛋白质的情况下,于 5%和 20%氧气条件下,在 EmbryoScope 延时培养箱中在七种培养基中培养 120 小时。

主要观察指标

分析七种培养基中 39 种单个氨基酸、有机酸、离子和元素的浓度。在 96 小时的培养后计算囊胚率和细胞周期时间,并进行了三次重复实验。

结果

在所分析的 39 种分析物中,葡萄糖、乳酸、丙酮酸、氨基酸、磷酸盐、钙和镁的浓度存在差异,这可能反映了对动物研究的不同解释。重要的微量元素,如铜和锌,未被检测到。一种培养基中胚胎无法发育,两种其他培养基中胚胎的发育受到氧气浓度和蛋白质存在的差异影响。

结论

培养基组成差异很大,特别是丙酮酸、乳酸和氨基酸差异显著。囊胚发育取决于培养基,并且与氧浓度和蛋白质存在存在相互作用。

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