Retinoic acid promotes expression of germline-specific genes in chicken blastoderm cells by stimulating Smad1/5 phosphorylation in a feeder-free culture system.

Publisher: BioMed Central Country of Publication: England NLM ID: 101088663 Publication Model: Electronic Cited Medium: Internet ISSN: 1472-6750 (Electronic) Linking ISSN: 14726750 NLM ISO Abbreviation: BMC Biotechnol Subsets: MEDLINE

Original Publication: [London] : BioMed Central, 2001-

Gene Expression Regulation, Developmental*/drug effects ; Gene Expression Regulation, Developmental*/physiology; Blastoderm/*cytology ; Blastoderm/*metabolism ; Germ Cells/*cytology ; Smad1 Protein/*metabolism ; Smad5 Protein/*metabolism ; Tretinoin/*pharmacology; Animals ; Batch Cell Culture Techniques ; Bioreactors ; Blastoderm/drug effects ; Cell Differentiation/drug effects ; Cell Differentiation/physiology ; Cells, Cultured ; Chick Embryo ; Chickens ; Dose-Response Relationship, Drug ; Germ Cells/metabolism ; Phosphorylation/drug effects

Background: Producing transgenic chickens with chicken blastodermal cells (cBCs) is inefficient due to the extremely low germline transmission capacity of cBCs. As chicken primordial germ cells (PGCs) have been reported as an efficient method for producing transgenic chickens, the inefficiency of cBCs could potentially be resolved by inducing them to differentiate into germ cells. However, whether chemical inducers are able to enhance cBCs germline competence in vitro is unknown and the molecular mechanisms of differentiation of chicken pluripotent cells into germ cells are poorly understood.
Results: We cultured cBCs with a monolayer morphology in E8 medium, a xeno- and feeder-free medium. We showed that retinoic acid (RA) treatment increased expression of germ cell-specific genes in cBCs. Using western blot, we determined that RA stimulated Smad1/5 phosphorylation. Moreover, Smad1/5 activation regulates the expression of germ cell-specific genes, as co-treatment with a Smad1/5 phosphorylation inhibitor or activator alters expression of these genes. We also demonstrate that Smad1/5 is required for RA-induced differentiation by RNA interference knockdown.
Conclusion: Our results demonstrated that E8 medium is able to maintain cBC growth for weeks and RA treatment induced germ cell differentiation of cBCs through the BMP-Smad1/5 signaling pathway.

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Keywords: Chicken blastodermal cells; E8 medium; Germ cell differentiation; Growth factor; Retinoic acid; Smad1/5 phosphorylation

0 (Smad1 Protein)
0 (Smad5 Protein)
5688UTC01R (Tretinoin)

Date Created: 20170222 Date Completed: 20170613 Latest Revision: 20231112

20240105

PMC5319176

10.1186/s12896-017-0332-y

28219352