A continuous culture of pluripotent fetal hepatocytes derived from the 8-day epiblast of the pig.
Continuous cultures of pluripotent parenchymal hepatocytes were derived from the epiblasts of 8-day-old pig blastocysts. The cells were polygonal and had phase-contrast dark, granular cytoplasm with prominent nuclei and nucleoli. These feeder-dependent cell cultures differentiated into large, multicellular, secretory, duct-like structures or formed small canaliculi between individual cells. Alternatively, the cells accumulated droplets that stained intensely with Oil Red O, a lipid-specific stain. Alpha-fetoprotein (AFP), albumin, and beta-fibrinogen mRNAs were expressed as the cells differentiated in culture. Serum-free medium that was conditioned by the cells contained transferrin, AFP, and albumin. The growth and viability of the cells were inhibited by transforming growth factor beta 1 (TGF beta 1) at concentrations > or = 1 ng/ml. The cell cultures grew slowly with doubling times of 2 to 3 d. One of the cultures, pig inner cell mass-19 (PICM-19), was passaged continuously for over 2 yr [> 100 population doublings (PD)] and appears to be an infinitely self-renewing cell population. The stem cell characteristics of the epiblast-derived fetal hepatocytes indicate that the cells may be unique for investigations of liver differentiation and organogenesis.
Talbot
N C
NC
U.S. Department of Agriculture, Beltsville Agricultural Research Center, Maryland 20705.
Rexroad
C E
CE
Powell
A M
AM
Pursel
V G
VG
Caperna
T J
TJ
Ogg
S L
SL
Nel
N D
ND
D000818:Animals; D015152:Blotting, Northern; D002454:Cell Differentiation; D002455:Cell Division; D002467:Cell Nucleus / Q000648:ultrastructure; D002478:Cells, Cultured; D003470:Culture Media; D017077:Culture Media, Conditioned; D003593:Cytoplasm / Q000648:ultrastructure; D005340:Fibrinogen / Q000235:genetics; D006168:Guinea Pigs; D015151:Immunoblotting; D008099:Liver / Q000196:embryology* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000378:metabolism; D012709:Serum Albumin / Q000235:genetics; D013552:Swine; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D000509:alpha-Fetoproteins / Q000235:genetics
D003470:Culture Media; D017077:Culture Media, Conditioned; D012333:RNA, Messenger; D012709:Serum Albumin; D016212:Transforming Growth Factor beta; D000509:alpha-Fetoproteins; D005340:Fibrinogen