Production of germline chimeras by transfer of chicken gonadal primordial germ cells maintained in vitro for an extended period.
We previously reported that germline chimeras could be produced by transfer of chicken gonadal primordial germ cells (gPGCs) cultured for a short term (5 days). This study was subsequently undertaken to examine whether gPGCs maintained in vitro for an extended period could retain their specific characteristics to induce germline transmission. Chicken (White Leghorn, WL) gPGCs were retrieved from embryos at stage 28 (5.5 days of incubation) and continuously cultured for 2 months in modified Dulbecco's minimal essential medium without subpassage and changing of the feeder cell layer. After the identification of gPGC characteristics using Periodic acid-Shiff's (PAS) reaction and anti stage-specific embryonic antigen-1 (SSEA-1) antibody staining at the end of the culture, cultured gPGCs were injected into the dorsal aorta of Korean Ogol Chicken (KOC) recipient embryos at stage 17 (2.5 days of incubation). Nineteen chickens (13 males and 6 females) were hatched, grown to sexual maturity, and subsequently subjected to testcross analysis employing artificial insemination with adult KOC. Of these, four (three males and one female) hatched chickens with white coat color. The percentage of germline chimerism was 21% (4/19). The results of this study demonstrated that gPGCs could maintain their specific characteristics for up to 2 months in vitro, resulting in the birth of germline chimeras following transfer to recipient embryos.
Han
Jae Yong
JY
School of Agricultural Biotechnology, Seoul National University, Suwon, South Korea. jaehan@snu.ac.kr
Park
Tae Sub
TS
Hong
Yeong Ho
YH
Jeong
Dong Kee
DK
Kim
Jin Nam
JN
Kim
Ki Dong
KD
Lim
Jeong Mook
JM
D000818:Animals; D030801:Animals, Genetically Modified; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics* / Q000502:physiology; D002678:Chimera / Q000235:genetics* / Q000502:physiology; D005260:Female; D005854:Germ Cells / Q000166:cytology / Q000502:physiology*; D006066:Gonads / Q000166:cytology / Q000502:physiology*; D008297:Male; D010858:Pigmentation / Q000235:genetics