Production of germ-line chimeras by transfer of cryopreserved gonadal primordial germ cells (gPGCs) in chicken
Production of germ-line chimeras by transfer of cryopreserved gonadal primordial germ cells (gPGCs) in chicken
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DOI：
10.1002/(sici)1097-010x(19980215)280:3
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发表时间：
1998-02-15
影响因子：
--
通讯作者：
Kuwana, T
中科院分区：
文献类型：
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作者：
Tajima, A;Naito, M;Kuwana, T
从5日龄白来宏(WL)胚胎的性腺原始生殖细胞(GPGCs)中采集生殖细胞。将收集的GPGCs悬浮在含有10%二甲基亚砜(DMSO)的冰冻液中。细胞悬浮液以1摄氏度/分钟的速度冷冻，直到温度达到-80摄氏度；然后将细胞浸入-196摄氏度的液氮中，储存长达3个月。将大约100个冷冻/解冻的gPGCs注射到每个被隔离的普利茅斯岩(BPR)胚胎的背部主动脉中，在注射生殖细胞之前从这些胚胎中提取血液。注射的胚胎被孵化直到孵化，孵化出的雏鸟被饲养到性成熟。当达到性成熟时，将受体雏鸡与正常的异性BPR交配进行子代测试。在来自4个gPGC受体的840只后代中，74只(8.8%)为表型白色。目前的结果表明，从鸡的性腺原基中采集的冷冻/解冻的gPGCs能够在鸡体内产生生殖系嵌合体。(C)1998年Wiley-Liss，Inc.
Gonadal primordial germ cells (gPGCs) were collected from gonadal anlage of 5-day-old White Leghorn (WL) embryos. Collected gPGCs were suspended in freezing medium containing 10% dimethyl sulphoxide (DMSO). The cell-suspension was frozen at 1 degrees C/min until the temperature reached -80 degrees C; cells were then immersed in liquid nitrogen at -196 degrees C and stored up to 3 mo. Approximately 100 frozen/thawed gPGCs were injected into the dorsal aorta of each Barred Plymouth Rock (BPR) embryo from which blood was drawn prior to germ-cell injection. The injected embryos were incubated until hatched, and hatched chicks were raised until sexually mature. Upon reaching sexual maturity, a progeny test was performed by mating recipient chicks with normal BPR of the opposite sex. Of 840 progeny produced from 4 gPGC recipients, 74 chicks (8.8%) were phenotypically white. Present results demonstrate that frozen/thawed gPGCs collected from gonadal anlage are capable of producing germ-line chimeras in chicken. (C) 1998 Wiley-Liss, Inc.