|
KMID : 0900919960200010043
|
|
Korean journal of Animal Reproduction
1996 Volume.20 No. 1 p.43 ~ p.52
|
|
|
In Vitro Culture Following Purfication of Mouse Spermatogenic Cells
|
|
Kim Jin-Hoi
Lee Sang-Min
Lee Hoon-Taek
|
|
|
Abstract
|
|
|
|
This study was carried out to establish the in vitro short-term culture system of developing male germ cells by purifing germ cells of various stages. The decapulated testicular cells were incubated with collagenase (lmg/ml) and try psin (2.5mg/ml) in HBSS. After separating male germ cell, the separated germ cells were stained with heamatoxylin/eosin and determined developing stages under light microscopy. The purity of pachtene spermatocytes a and round spermatid were 85%, respectively. Yield of total male germ cells was highly variable between individuals, with a mean value of 3.5 to 4.5 {times} 10^7 cells/testis. Viability of the cell was over 97% after separation. In DMEM medium, the optimal cell number for culture is approximately 1 x 10^5 cells/dish, but low cell den-sities than 1 {times} 10^5 cell/dish showed a decreased cell viability. Furthermore, about :36.8% of pac-hytene cells was successfully cultured for 6 days and some of cells were developed to secondary spermatids and round spermatids. Therefore, our data suggested that this culture conditions will be utilize as a feasible tools to produce tran-sgenic livestock using techniques such as intrac-ytoplasmic injection and cell fusion.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
|
|