title: Delayed BMP4 exposure increases germ cell differentiation in mouse embryonic stem cells.
abstract: Fate mapping studies have revealed that bone morphogenetic protein 4 (BMP4) signaling has a key role in segregation of primordial germ cells from proximal epiblast. Adding BMP4 to the culture media of embryonic stem (ES) cells could induce expression of germ cell markers; however, to provide a desired number of germ cells has remained a challenge. In the current study, we intended to establish an in vitro system to obtain reliable germ cells derived from ES cells. Differentiation was induced in ES cells via embryoid body (EB) and monolayer culture system. Cells were cultured with BMP4 from the beginning (++BMP4) or after 48 hours (+BMP4) of culturing for five days. The cultures were assessed for alkaline phosphatase (ALP) activity, expression of Oct4, Mvh and c-kit. In EB culture protocol, the expression of Mvh, Oct4 and ALP activity significantly increased in +BMP4 culture condition, but a significant down-regulation in the expression of germ cell markers was shown in ++BMP4 condition compared with the control group. Parallel differentiation experiments using monolayer culture system indicated the number of putative germ cells did not change. In the current study, we compared two differentiation methods (EB and monolayer) to achieve an optimal germ cell production. The EBs with a short exposure time period to BMP4, showing typical characteristics of germ cells. Therefore, our approach provides a strategy for the production of germline cells from ES cells.
authors: Department of Anatomical Sciences, School of Medicine, Shiraz, Iran; bahmans@sums.ac.ir.
authors: Tahereh
authors: T
authors: Talaei-Khozani
authors: Nehleh
authors: N
authors: Zarei Fard
authors: Soghra
authors: S
authors: Bahmanpour
authors: Mansoureh
authors: M
authors: Jaberipour
authors: Ahmah
authors: A
authors: Hosseini
authors: Tahereh
authors: T
authors: Esmaeilpour
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D005718:Gametogenesis / Q000187:drug effects; D005854:Germ Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D013997:Time Factors
chemical_list: C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4