Directed differentiation of chick embryonic germ cells into neural cells using retinoic acid induction in vitro.
Embryonic germ cells (EG) are undifferentiated stem cells isolated from cultured primordial germ cells (PGC). Like embryonic stem cells (ES), EG are also capable of proliferation and self-renewal and have the capacity to differentiate in vitro into all cell types. To date, it has been proven that ES are capable of directed differentiation into neural precursors and progenies in mammals. However, similar studies on EG in mammals and other species are few. This investigation aimed to induce chick EG to differentiate into neural cells and compare the difference of efficiency between directed differentiation and spontaneous differentiation. EG were isolated and identified from 5.5-day chick gonadal PGC, incubated and passaged in conditioned medium. After the formation of embryoid bodies (EB), EB were grown in suspension and induced by retinoic acid (RA), using a protocol named 4-/4+, to make the formation of neurospheres and progenies. RT-PCR and immunocytochemistry analysis demonstrated that neural-specific markers can be detected after directed induction. Moreover, EG differentiated into neural lineage cells using 4-/4+ protocol much more efficiently than that in the spontaneous differentiation with fluorescence-activated cell sorting (FACS) analysis. The results revealed that RA can obviously promote the directed differentiation of chick EG into neural lineage.
Wang
Juan
J
Department of Biotechnology, Binzhou Medical College, 264003 Yantai, Shandong Province, PR China.
Jiao
Fei
F
Pan
Xiao-Hong
XH
Xie
Shu-Yang
SY
Li
Zun-Ling
ZL
Niu
Xin-Hua
XH
Du
Li-Xin
LX
D000818:Animals; D019521:Body Patterning; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005854:Germ Cells / Q000187:drug effects*; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D064231:Nestin; D009457:Neuroglia / Q000187:drug effects; D009474:Neurons / Q000502:physiology*; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques; D014212:Tretinoin / Q000494:pharmacology*
D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; D064231:Nestin; D055748:SOXB1 Transcription Factors; D014212:Tretinoin