Induced Multilineage Differentiation of Chicken Embryonic Germ Cells via Embryoid Body Formation

Abstract

Although the pluripotent and proliferative capacity of embryonic germ (EG) cells is thought to be equivalent to that of embryonic stem (ES) cells, there has been far less attention focused on the potential use of EG cells for applications in developing novel strategies of tissue transplantation in the treatment of degenerative diseases. In this study, EG cells were derived from primordial germ cells (PGCs) of genital ridges of 4-day-old chicken embryos. These cells satisfied the criteria previously used for defining chicken EG cells by using the expression of markers characteristic to ES cells. When injected subcutaneously, chicken EG cells could form teratomas that enable differentiation into a wide range of tissue types of all three primary cell lineages including neural cells, cartilage, forming bone, adipocytes, blood vessels, smooth muscle, and secretory epithelia in the recipients. Furthermore, cells in embryoid bodies (EBs) expressed lineage-specific markers of three germ layers and could be induced to differentiate into more advanced stages of various committed cell types, including dopamine and cholinergic neurons, astrocytes, oligodendrocytes, adipocytes, and hepatocytes, which were demonstrated by immunocytochemical staining or RT-PCR analysis. These findings support the multilineage differentiation capability of chicken pluripotent EG cells, thus confirming the presumption that chicken embryos may be used as a potential model for better understanding the mechanisms of tissue-specific differentiation and regeneration that will help to devise strategies based on the transplantation of stem cell-derived tissues for restoring function to damaged or diseased tissues.

Effect of Dynamic DNA Methylation and Histone Acetylation oncPouVExpression in Differentiation of Chick Embryonic Germ Cells

“…For alkaline phosphatase (AP) staining, fixed cells were immersed in a filtered AP staining solution [2 mg naphtol AS-MX phosphate, 200 mL N,N-dimethylformamide, 9.8 mL of 0.1 M Tris (pH 8.2), and 10 mg Fast Red TR salt] for 30 min, and then rinsed three times with 1 · PBS. To detect expression of stem cell markers, EG cell colonies were examined for expression of the stage-specific embryonic antigen-1 (SSEA-1), and SSEA-3 by reaction with streptavidin-horseradish peroxidase, 3,3¢-diaminobenzidine, and hydrogen peroxide (Sigma) via the biotinylated antimouse secondary antibody (1:150; Chemicon) [21].…”

Retinoic acid promotes proliferation of chicken primordial germ cells via activation of PI3K/Akt‐mediated NF‐κB signalling cascade

“…Primary culture was done as in a previous study (Wu et al, 2010). The gonads were carefully dissected from the embryonic mesonephros with a fine glass needle under a stereomicroscope (SZX16, Olympus).…”

Livestock Induced Pluripotent Stem Cells

“…2010). Previous publications reported the differentiation of chicken embryonic germ cells (EGs) into multiple lineages, indicating that these cells may be utilized to perform transplantation studies in birds (Wu et al. 2010).…”

“…Avian species are relatively small in size and the embryo can be easily accessed for manipulation enabling cells or tissues to be transplanted, which is not possible in mammals (Kulesa et al 2010). Previous publications reported the differentiation of chicken embryonic germ cells (EGs) into multiple lineages, indicating that these cells may be utilized to perform transplantation studies in birds (Wu et al 2010). Quail iPSCs are easily maintained in feederfree and chemical-defined culture systems, and form EBs and demonstrate differentiation into all three germ layers: TUJ1 (ectoderm), SOX17 (endoderm) and alpha smooth muscle actin (aSMA, mesoderm).…”