Abstract
Chicken embryonic stem (ES) cells are useful for producing transgenic chickens and preserving genetic material in avian species. In this study, the differentiation potential of chicken ES cells was investigated in vitro . Chicken ES cells were differentiated into osteoblasts cultured for 15 to 21 days in the induction media containing dexamethasone (DEX), β-glycerol phosphate (β-GP) and/or vitamin C (Vit C) respectively and differentiation rates ranging from 40 to 72% confirmed by Von Kossa, cytochemical and immunohistochemical staining. Chicken ES differentiated into neuron-like cells cultured for 3 to 7 days in the induction media containing retinoic acid (RA) and 3-isobutyl-1-methylxanthine (IBMX) and differentiation rates ranging from 70 to 84% were identified by toluidine blue staining and immunohistochemical staining. Also chicken ES was differentiated into adipocytes cultured for 21 days in the induction media containing DEX, insulin (Ins) and/or IBMX and differentiation rates ranging from 74 to 91% identified by oil red-O and reverse transcriptase-polymerase chain reaction (RT-PCR) for peroxisome proliferator activated receptor-γ (PPAR-γ) gene expression. These data suggest that like mammalian ES, chicken ES can differentiate into different cell types in vitro . Key words: Chicken embryonic stem cells, in vitro, directional differentiation, osteoblasts, neuron-like cells, adipocytes.
Highlights
Embryonic stem (ES) cells are derived from the inner cell mass (ICM) of mammalian preimplantation blastula or primordial germ cells (PGCs) and cultured to establish cell clone lines in vitro (Xiao et al, 2000)
Chicken embryonic stem (ES) cells were differentiated into osteoblasts cultured for 15 to 21 days in the induction media containing dexamethasone (DEX), β-glycerol phosphate (β-GP) and/or vitamin C (Vit C) respectively and differentiation rates ranging from 40 to 72% confirmed by Von Kossa, cytochemical and immunohistochemical staining
Chicken ES was differentiated into adipocytes cultured for 21 days in the induction media containing DEX, insulin (Ins) and/or IBMX and differentiation rates ranging from 74 to 91% identified by oil red-O and reverse transcriptase-polymerase chain reaction (RT-PCR) for peroxisome proliferator activated receptor-γ (PPAR-γ) gene expression
Summary
Embryonic stem (ES) cells are derived from the inner cell mass (ICM) of mammalian preimplantation blastula or primordial germ cells (PGCs) and cultured to establish cell clone lines in vitro (Xiao et al, 2000). Cells can differentiate into multiple somatic cell types in vitro, showing a new avenue for cell-replacement therapy. Yagami et al (2004) demonstrated that mesenchymal stem cells derived from human chondrogenic cell line could differentiate into adipocytes. As an optimal origin of stem cell, reports on in vitro differentiation of chicken ES cells are limited. We investigated the multipotential differentiation of chicken ES cells into different cell types in vitro. The experimental data showed that high percentages of chicken ES cells could differen-
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