补充凝集素对水牛卵母细胞体外成熟的改善及Cx43、GDF-9、FGF-4和纤连蛋白mRNA转录本的表达分析
Improvement of in vitro oocyte maturation with lectin supplementation and expression analysis of Cx43, GDF-9, FGF-4 and Fibronectin mRNA transcripts in Buffalo (Bubalus bubalis).
作者信息
Pandey Alok, Gupta Neelam, Gupta S C
机构信息
Transgenic Research Laboratory, DNA Finger Printing Unit, National Bureau of Animal Genetic Resources, Karnal 132001, India.
出版信息
J Assist Reprod Genet. 2009 Jun;26(6):365-71. doi: 10.1007/s10815-009-9314-x. Epub 2009 Jul 21.
PURPOSE
To optimize In vitro maturation (IVM) of quality oocytes for embryo production through IVF and SCNT.
METHODS
Buffalo oocytes were in vitro matured in the presence of the pokeweed lectin (Phytolacca americana), a potent lymphocyte mitogen. Lectin was supplemented in TCM + 10% FBS at the doses of 0, 1, 5, 10, 15, 20 and 40 microg/ml and cumulus expansion and gene expression patterns were characterized.
RESULTS
The degree of cumulus expansion in different lectin treatment levels improved from 1.1 at 1 Ag/ml level to 3.60 at 10 microg/ml level and then decreased in higher concentration 20 microg/ml (1.66) and 40 microg/ml (0.64). IVF embryos showed highest cleavage rate (88.8%) in 10 microg/ml lectin treatment. Expression of all mRNA transcript studied (Cx43, GDF 9, FGF-4 and Fibronectin) was positively correlated with cumulus expansion and polar body extrusion.
CONCLUSIONS
Mitogenic lectin supplemented maturation media improves oocyte quality for in vitro embryo production.
目的
通过体外受精（IVF）和体细胞核移植（SCNT）优化优质卵母细胞的体外成熟（IVM），以用于胚胎生产。
方法
将水牛卵母细胞在商陆凝集素（美洲商陆）存在的情况下进行体外成熟培养，商陆凝集素是一种有效的淋巴细胞促有丝分裂原。在含有10%胎牛血清（FBS）的组织培养液（TCM）中添加不同剂量（0、1、5、10、15、20和40微克/毫升）的凝集素，并对卵丘扩展和基因表达模式进行表征。
结果
不同凝集素处理水平下的卵丘扩展程度从1微克/毫升水平时的1.1提高到10微克/毫升水平时的3.60，然后在更高浓度（20微克/毫升时为1.66，40微克/毫升时为0.64）下降低。在10微克/毫升凝集素处理下，体外受精胚胎显示出最高的卵裂率（88.8%）。所研究的所有mRNA转录本（Cx43、生长分化因子9、成纤维细胞生长因子-4和纤连蛋白）的表达与卵丘扩展和极体排出呈正相关。
结论
添加有促有丝分裂凝集素的成熟培养液可提高用于体外胚胎生产的卵母细胞质量。
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