摘要
旨在对影响鸡胚胎原始生殖细胞(PGC s)体外培养、传代过程的各种影响因素进行探讨。将分离到的19期鸡胚胎生殖腺的PGC s在以鸡胚成纤维细胞(CEF)为饲养层,添加细胞因子的培养体系中进行培养、传代。经冷冻保存的PGC s在添加细胞因子的培养体系中能增殖,并具有分化能力。从细胞形态、集落形态、增殖生长能力、核型检测、碱性磷酸酶测定等方面对培养过程中的鸡胚胎PGC s进行检测。结果表明,采用传至3代的CEF为饲养层,多次离散法进行传代所获得的5代鸡胚胎PGC s不但能有效维持PGC s的未分化状态和正常二倍体核型,也具有其作为多能性胚胎干细胞的一系列特征。
This study was performed to characterize some related effects during chicken PGCs culture. Chicken PGCs were isolated from stage 19 genital ridge cultured on chicken fibroblast feeder layer in presence of growth factors. Frozen-thawed PGCs continued to proliferated when growth factors was added to the culture medium. However,differentiation into fibroblastlike cells or neuronal cells were observed after 1 or 2 passages. The PGCs in the culture were monitored according to their morphology,proliferative capability and karyotypes,and the alkaline phosphatase activity. The results suggested that it＇s a reasonable method to dissociate PGCs colonies using ＂several times dissociation method＂ after 6 days proliferation,the 5th passage PGCs that have been cultured in vitro not only maintained the undifferentiated state and normal diploid karyotype,but also bore the necessary characteristics of chicken PGCs,thus providing a promising source of donor-karyoplasts for gene transfer proced u res.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2006年第4期444-447,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30371031)
