Manipulation of blastodermal cells.
Blastodermal cells isolated from newly laid, unincubated eggs are virtually uncommitted cells that exhibit many of the properties of pluripotential stem cells. They can be transferred from donor to recipient embryos and contribute to both somatic tissues and the germline. Blastodermal cells that have been maintained in culture for 7 d express the epitopes ECMA-7 and SSEA-1, which are also expressed by mouse embryonic stem cells. After culture for up to at least 7 d, blastodermal cells retain the ability to differentiate into somatic tissues and the germline both in vivo and in vitro. Proliferation in the absence of differentiation of blastodermal cells is stimulated by the presence of Leukemia Inhibitory Factor (LIF) and other ligands that interact with the gp130 receptor, and differentiation is stimulated by exposure to retinoic acid. Blastodermal cells also possess high levels of telomerase activity, which is shared by immortalized cells and cells within the germline. Blastodermal cells can be transfected and will express foreign genes both in vivo and in vitro. Transfected cells can be isolated by fluorescence activated cell sorting and can be cryopreserved without losing their ability to contribute to either somatic tissues or the germline. These properties of blastodermal cells make them ideal vectors for introducing genetic modifications to the germline.
Etches
R J
RJ
Department of Animal and Poultry Science, University of Guelph, Ontario, Canada.
Clark
M E
ME
Zajchowski
L
L
Speksnijder
G
G
Verrinder Gibbins
A M
AM
Kino
K
K
Pain
B
B
Samarut
J
J
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001756:Blastoderm / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology* / Q000254:growth & development / Q000502:physiology; D002678:Chimera; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary; D005822:Genetic Vectors; D006131:Growth Inhibitors / Q000032:analysis; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000032:analysis; D051379:Mice; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D013997:Time Factors; D014162:Transfection
D006131:Growth Inhibitors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D000469:Alkaline Phosphatase