摘要
本研究以版纳微型猪近交系成年母猪为材料,采用冷胰酶消化法获得成年猪成纤维细胞后,进行细胞的生物学特征检测,研究其作为供体细胞对体细胞核移植效果的影响。结果表明,版纳微型猪近交系成年猪的成纤维细胞呈典型的成纤维细胞形态,细胞冻存前和复苏后存活率分别为97.2%和92.6%(P>0.05),生长曲线呈"S"形,细胞群体倍增时间为36h,体外培养14代后仍能保持正常核型。以其作为核移植供体细胞,核移植重构胚卵裂率、囊胚率及囊胚细胞数分别为61.9%、13.0%和37枚,移植了重构胚的3头代孕母猪均妊娠,并且有1头母猪产下1头正常的克隆猪。综上所述,利用版纳微型猪近交系成年猪能建立稳定的成纤维细胞系,该细胞系作为核移植供体细胞可获得版纳微型猪近交系克隆猪。
Fibroblast cells were isolated from adult Banna Miniature Inbred pig with cold trypsin digestion method and subcultured. Through study of cell biology characteristics, effects of fibro- blast cells as the donor cells on somatic cell nuclear transfer was investigated. The results showed that isolated fibroblast cells were morphologically consistent with typical fibroblast, cell viability was 97.20/00 before freezing and 92.6~//oo after thawing （P^0.05）. The cell growth curve was ＇$~ shape and population doubling time was approximately 36 h. The karyotype was normal after 14 passages of subculture in vitro. When isolated fibroblast cells were used as donor cells for somat- ic cell nuclear transfer, the cleavage rate, the blastocyst rate and cell number of blastocyst were 61.9~, 13.0~ and 37, respectively. One of three surrogate pigs after somatic cell nuclear trans- fer with fibroblast cells as donor cells was pregnant and gave birth to one clone pig. In brief, fi- broblast cell line could be established from adult Banna Miniature Inbred pig. The isolated fibro-blast cells could be used as donor cells for somatic cell nuclear transfer and gave birth to the clone pig of Banna Miniature Inbred pig.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2012年第9期1491-1498,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金项目(31060308)
973计划项目(2011CB944200)
