Generation of Male Germ Cells from Embryonic Stem Cells

Background: Artificial gamete formation accompanied with progressive success in mouse and human. Stem cell therapy can be served as an alternative therapy for male infertility via formation of male gametes. Here we differentiated mouse Embryonic stem cells (ESc) in to male germ cells using Embryoid body (EB) technique and in the presence of appropriate inducers. Methods: Primordial germ cells (PGCs), were induced followed the exposure of EBs to various concentrations of BMP4 for 4 and 7 days culture. The specific germ cell markers were detected using Real Time PCR, Immunocytochemistry and flow cytometry. PGCs cells were cultured in mouse spermatogonial stem cell medium, containing: DMEM, μM β-mercaptoethanol, Lglutamine, Bovine Serum, Albumin, transferrin, insulin, bFGF and GDNF. The specific spermatogonial lineage markers were detected using RT- PCR, Immunocytochemistry techniques. Results: Our results proved Mvh marker was expressed in the adge of induced EBs. The highestexpression of gene Oct4 gene in the untreated group and it reduced in the presence with BMP4. The expression of Stella and Mvh as a pre and post migratory markers exhibited the most expression in experiment group. The expression of specific spermatogonial markers DAZL, PLZF, UTF1, VASA were confirmed via RT- PCR and Immunocytochemistry methods. Conclusion: It is conclude BMP4 is an effective factor to induction of PGCs from mouse ESC. Our culture system leads to derivation of Germ cells lineages in EB population and increase the differentiation of ESCs into spermatogenic cells.

کلیدواژه ها:

Artificial gamete ، Embryoid Stem Cells ، Primordial germ cells ، BMP4 ، Embryoid Bodies ، Mvhpositive cells