Establishment of new murine embryonic stem cell lines for the generation of mouse models of human genetic diseases.
Embryonic stem cells are totipotent cells derived from the inner cell mass of blastocysts. Recently, the development of appropriate culture conditions for the differentiation of these cells into specific cell types has permitted their use as potential therapeutic agents for several diseases. In addition, manipulation of their genome in vitro allows the creation of animal models of human genetic diseases and for the study of gene function in vivo. We report the establishment of new lines of murine embryonic stem cells from preimplantation stage embryos of 129/Sv mice. Most of these cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines obtained were analyzed on the basis of their alkaline phosphatase activity and their capacity to form complex embryoid bodies with rhythmically contracting cardiomyocytes. Two lines, USP-1 and USP-3, with the best in vitro characteristics of pluripotency were used in chimera-generating experiments. The capacity to contribute to the germ line was demonstrated by the USP-1 cell line. This cell line is currently being used to generate mouse models of human diseases.
Sukoyan
M A
MA
Departamento de Biologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão 277, 05508-900 São Paulo, Brazil.
Kerkis
A Y
AY
Mello
M R B
MR
Kerkis
I E
IE
Visintin
J A
JA
Pereira
L V
LV
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line / Q000166:cytology* / Q000201:enzymology; D002678:Chimera; D004195:Disease Models, Animal; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D030342:Genetic Diseases, Inborn / Q000235:genetics* / Q000473:pathology; D005854:Germ Cells; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D013234:Stem Cells / Q000166:cytology* / Q000201:enzymology
D000469:Alkaline Phosphatase