Bone morphogenetic protein 4 mediates human embryonic germ cell derivation.
Human primordial germ cells (PGCs) have proven to be a source of pluripotent stem cells called embryonic germ cells (EGCs). Unlike embryonic stem cells, virtually little is known regarding the factors that regulate EGC survival and maintenance. In mice, the growth factor bone morphogenetic protein 4 (BMP4) has been shown to be required for maintaining mouse embryonic stem cells, and disruptions in this gene lead to defects in mouse PGC specification. Here, we sought to determine whether recombinant human BMP4 could influence EGC derivation and/or human PGC survival. We found that the addition of recombinant BMP4 increased the number of human PGCs after 1 week of culture in a dose-responsive manner. The efficiency of EGC derivation and maintenance in culture was also enhanced by the presence of recombinant BMP4 based on alkaline phosphatase and OCT4 staining. In addition, an antagonist of the BMP4 pathway, Noggin, decreased PGC proliferation and led to an increase in cystic embryoid body formation. Quantitative real-time (qRT)-polymerase chain reaction analyses and immunostaining confirmed that the constituents of the BMP4 pathway were upregulated in EGCs versus PGCs. Downstream activators of the BMP4 pathway such as ID1 and phosphorylated SMADs 1 and 5 were also expressed, suggesting a role of this growth factor in EGC pluripotency.
Hiller
Marc
M
Department of Gynecology and Obstetrics, Institute for Cellular Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Liu
Cyndi
C
Blumenthal
Paul D
PD
Gearhart
John D
JD
Kerr
Candace L
CL
D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology* / Q000502:physiology; D052004:Bone Morphogenetic Protein Receptors / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D058732:Embryoid Bodies / Q000187:drug effects; D005260:Female; D020869:Gene Expression Profiling; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects; D005865:Gestational Age; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D011247:Pregnancy; D011994:Recombinant Proteins / Q000494:pharmacology*; D051785:Smad Proteins / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
D000943:Antigens, Differentiation; D055415:Bone Morphogenetic Protein 4; D011994:Recombinant Proteins; D051785:Smad Proteins; D052004:Bone Morphogenetic Protein Receptors