Development of avian primordial germ cells in vivo and in vitro
Başlık çevirisi mevcut değil.
- Tez No: 400619
- Danışmanlar: DR. JAMES N. PETITTE
- Tez Türü: Doktora
- Konular: Veteriner Hekimliği, Zooloji, Veterinary Medicine, Zoology
- Anahtar Kelimeler: primordial germ cells, chicken, embryo, SSEA-1, basic fibroblast growth factor, stem cell factor, brachyury)
- Yıl: 1998
- Dil: İngilizce
- Üniversite: North Carolina State University
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 212
Özet
Özet yok.
Özet (Çeviri)
The temporal and spatial patterns of the origin of chick primordial germ cells (PGCs) between stages IX - XIV were investigated using immunohistological analysis of whole mount embryos and histological sections with a monoclonal antibody (MC-480) that recognizes Stage Specific Embryonic Antigen-1 (SSEA-1), and using the culture of dispersed blastoderms at stages IX - XIV with and without STO feeder cells, a SIM mouse embryo-derived, Thioguanine- and Oubain-resistant fibroblast cell line. The carbohydrate epitope recognized by MC-480 first appeared at stage X on cells in association with polyingressing cells on the ventral surface of the epiblast and later on the dorsal surface of the hypoblast. The SSEA-1-positive hypoblast cells gave rise to chicken PGCs when injected into the sub-germinal cavity of quail blastoderms or when cultured on a feeder-layer of quail blastodermal cells. When individual stage IX-XIV embryos were dispersed and cultured without a feeder-layer, 25-45 PGCs/embryo were detected using stages X-XIV, but not with stage IX blastoderms. Similar results were obtained when blastodermal cells were cultured either on gelatin-coated tissue-culture plastic or on feeder-layers of CV-1 cells. However, the culture of blastodermal cells obtained from the area pellucidae of stages IX-Xlll using feeder-layers of STO cells yielded about 110-170 PGCs/embryo. The addition of STO cell-conditioned medium (STO-CM) to blastodermal cells cultured either on gelatin-coated plastic or on feeder-layers of CV-1 cells resulted in a significant increase in the number of PGCs compared to cultures carried out without STO-CM. Blastodermal cells cultured on feeder-layers of CV-1 cells in the presence of recombinant avian stem cell factor (SCF) (100 ng/mL) yielded a significantly higher number of PGCs after 48 hr of culture compared to those cultured in the absence of SCF. Conversely, the supplementation of blastodermal cell cultures carried out on feeder-layers of either STO or CV-1 cells with basic fibroblast growth factor (bFGF) (10 and 100 ng/ mL) significantly reduced the number of PGCs. The inhibitory effect of bFGF on the culture of PGCs on STO feeder-layers was associated with the down-regulation of SSEA-1 and was concomitant with the up-regulation of brachyury, a general marker for early mesodermal cells. In accordance with the possible involvement of SCF in the development of avian PGCs, mRNA transcripts for SCF and its tyrosine kinase receptor, c-kit, were detected at pre-streak stages of development. In situ hybridization analyses demonstrated that SCF was expressed in epithelial cells forming the gonadal ridge, the final destination of PGCs, while c-kit was expressed by PGCs themselves. On the basis of these observations, it is proposed that 1) the segregation and development of avian PGCs is a gradual process associated with the translocation of SSEA-1-positive cells from the ventral surface of the area peliucida at stage X to the dorsal side of the hypoblast at stages XI-XIV and 2) the differentiation of avian PGCs during the formation of area peltucida is not a cell autonomous process and is the result of specific local cell-cell interactions which might be mediated by peptide growth factors and their corresponding receptors, such as the SCF/c-kit ligand-receptor complex. (
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