Pluripotency of bank vole embryonic cells depends on FGF2 and activin A signaling pathways.
The objective of this study was to investigate the capability of bank vole (Myodes glareolus) embryonic cells to sustain their pluripotent character during in vitro culture, and to determine the optimal conditions for derivation of embryonic stem (ES) cells. We compared the presence of specific pluripotency (Oct4, Ssea1) and differentiation markers (Gata4 - primitive endoderm marker; Cdx2 - trophectoderm marker) in blastocysts and inner cell mass (ICM) outgrowths obtained from blastocysts of bank vole, and two mouse hybrids F1(C57Bl/6xCBA/H) and F1(C57Bl/6x129/Sv), which differ in the permissiveness of giving rise to ES cells. We found that, in contrast to mouse, the expression of pluripotency markers in the cells of bank vole ICM outgrowths is progressively downregulated and rapidly lost by the 4th day of culture. This correlates with the appearance of cells expressing Gata4 and Cdx2, indicating differentiation towards primitive endoderm and derivatives of trophectoderm, respectively. We have also shown that heterologous cytokine leukaemia inhibitory factor (LIF) in conjunction with either homologous or heterologous feeder layer is unable to delay differentiation and preserve pluripotency of bank vole embryonic cells. Thus, the conditions optimised for mouse do not support the maintenance of bank vole embryonic cells in the undifferentiated state and do not allow for the isolation of the ES cells. Instead, combination of fibroblast growth factor 2 and activin A allows retention of Oct4 expression in bank vole blastocyst outgrowths during 4-day culture, indicating that signaling pathways operating in human, rather than mouse ES cells, might be involved in the process of self-renewal of bank vole embryonic cells.
Suwinska
Aneta
A
Department of Embryology, Institute of Zoology, University of Warsaw, Warsaw, Poland.
Tarkowski
Andrzej K
AK
Ciemerych
Maria A
MA
D028341:Activins / Q000378:metabolism*; D000818:Animals; D003411:Arvicolinae / Q000196:embryology; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL / Q000196:embryology; D008808:Mice, Inbred CBA / Q000196:embryology; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D015398:Signal Transduction
D053605:Leukemia Inhibitory Factor; C422405:activin A; D016222:Fibroblast Growth Factor 2; D028341:Activins