Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 1 de 1
Filter
Add more filters










Database
Language
Publication year range
1.
Hum Reprod ; 25(1): 168-78, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19840987

ABSTRACT

BACKGROUND: Signaling mechanisms involved in early human germ cell development are largely unknown and believed to be similar to mouse germ cell development; however, there may be species specific differences. KIT ligand (KITL) and Bone morphogenetic protein 4 (BMP4) are necessary in mouse germ cell development and may play an important role in human germ cell development. METHODS: KITL signaling studies were conducted by differentiating human embryonic stem cells (hESCs) on KITL wild-type, hetero- or homozygous knockout feeders for 10 days, and the effects of BMP signaling was determined by differentiation in the presence of BMP4 or its antagonist, Noggin. The formation of germ-like cells was ascertained by immunocytochemistry, flow cytometry and quantitative RT-PCR for germ cell markers. RESULTS: The loss of KITL in enrichment and differentiation cultures resulted in significant down-regulation of germ cell genes and a 70.5% decrease in germ-like (DDX4+ POU5F1+) cells, indicating that KITL is involved in human germ cell development. Moreover, endogenous BMP signaling caused germ-like (DDX4+ POU5F1+) cell differentiation, and the inhibition of this pathway caused a significant decrease in germ cell gene expression and in the number of DDX4+ POU5F1+ cells. Further, we demonstrated that eliminating feeders but maintaining their secreted extracellular matrix is sufficient to sustain the increased numbers of DDX4+ POU5F1+ cells in culture. However, this resulted in decreased germ cell gene expression. CONCLUSIONS: From these studies, we establish that KITL and BMP4 germ cell signaling affects in vitro formation of hESC derived germ-like cells and we suggest that they may play an important role in normal human germ cell development.


Subject(s)
Bone Morphogenetic Protein 4/pharmacology , Cell Differentiation , Embryonic Stem Cells/drug effects , Germ Cells/cytology , Stem Cell Factor/pharmacology , Animals , Bone Morphogenetic Protein 4/antagonists & inhibitors , Carrier Proteins/pharmacology , Coculture Techniques , DEAD-box RNA Helicases/metabolism , DNA Methylation , Embryonic Stem Cells/cytology , Genome, Human , Germ Cells/growth & development , Germ Cells/metabolism , Humans , Mice , Octamer Transcription Factor-3/metabolism , Signal Transduction
SELECTION OF CITATIONS
SEARCH DETAIL
...