Embryonic stem cell-like cells established by culture of adult ovarian cells in mice.
OBJECTIVE
To suggest an alternative strategy for deriving histocompatible stems cells without undertaking genetic manipulation.


METHODS
Prospective approach using an animal model.


METHODS
Stem cell and bioevaluation laboratory, Seoul National University.


METHODS
F1 (C57BL6 X DBA2) and outbred (ICR) mice.


METHODS
Ovarian stroma cells of less than 40 mum in diameter were subcultured with fibroblast monolayer, and colony-forming cells were characterized.


METHODS
Stemness, genotype, and imprinted gene methylation.


RESULTS
Two-lines of colony-forming cells were established, which expressed markers specific for embryonic stem cells (ESC) and formed embryoid bodies and teratomas. Complete matching of microsatellite markers with the cell donor strain confirmed their establishment from ovarian tissue, and identification of both homozygotic and heterozygotic chromosomes raised the possibility of their derivation from parthenogenetic oocytes. However, the use of cells smaller than mature oocytes for primary culture, the difference in imprinted gene methylation compared with parthenogenetic ESCs, and failure to establish the ESC-like cells by primary follicle culture collectively suggested the irrelevancy to gametes.


CONCLUSIONS
Coculture of adult ovarian cells with somatic fibroblasts can yield colony-forming cells having ESC-like activity, which may provide an alternative for establishing autologous stem cells from adults that can be obtained without genetic manipulation.
Gong
Seung Pyo
SP
Department of Agricultural Biotechnology, Seoul National University, Seoul, South Korea.
Lee
Seung Tae
ST
Lee
Eun Ju
EJ
Kim
Dae Yong
DY
Lee
Gene
G
Chi
Sung Gil
SG
Ryu
Byung-Kyu
BK
Lee
Chae Hyun
CH
Yum
Kyung Eun
KE
Lee
Ho-Joon
HJ
Han
Jae Yong
JY
Tilly
Jonathan L
JL
Lim
Jeong Mook
JM
D000818:Animals; D018920:Coculture Techniques; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000096:biosynthesis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D018398:Homeodomain Proteins / Q000096:biosynthesis; D051379:Mice; D008813:Mice, Inbred ICR; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D006080:Ovarian Follicle / Q000166:cytology*; D010312:Parthenogenesis; D013234:Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology; D019098:Telomerase / Q000096:biosynthesis; D014157:Transcription Factors / Q000096:biosynthesis; D000097584:DNA Methyltransferase 3B
D004248:DNA (Cytosine-5-)-Methyltransferases; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D019098:Telomerase; D014157:Transcription Factors; D000097584:DNA Methyltransferase 3B; C475540:Nanog protein, mouse; C513672:Zfp42 protein, mouse; C509187:Tert protein, mouse