Directional differentiation of chicken spermatogonial stem cells in vitro

被引:19
作者
Li, Bichun [1 ]
Wang, X.-Y. [1 ]
Tian, Zhiquan [1 ]
Xiao, X.-J. [1 ]
Xu, Q. [1 ]
Wei, C.-X. [1 ]
Y, F. [1 ]
Sun, H.-C. [1 ]
Chen, G.-H. [1 ]
机构
[1] Yangzhou Univ, Coll Anim Sci & Technol, Yangzhou, Peoples R China
基金
中国国家自然科学基金;
关键词
adipocytes; chicken spermatogonial stem cells; in vitro differentiation; neuron-like cells; osteoblasts; OSTEOBLASTS; GENERATION; ACID;
D O I
10.3109/14653240903518155
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background. Mammalian spermatogonial stem cells (SSC) are able to differentiate into different cell types in vitro, which are valuable sources for regenerative medicine and gene transfer studies. We investigated the differentiation potential of chicken SSC into osteoblasts, neuron-like cells and adipocytes in vitro. Methods. Chicken SSC from the testes of 18- and 20-day-old chicken embryos were cultured in different induction media for three passages in vitro. For differentiation into osteoblasts, SSC were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 1x10<SU-4</SU mu mol/mL desamethasone, 10 mu mol/mL (beta-sodium glycerophosphate and 0.05 mg/mL vitamin C, and examined by microscopy after Von Kossa's, cytochemical and immunohistochemical staining. For differentiation into neuron-like cells, SSC were cultured in DMEM supplemented with 1 x 10<SU-3</SU mu mol/mL retinoic acid (RA), 5.0 mu mol/mL 3-isobutyl-1-methylxanthine (IBMX) and examined by microscopy after toluidine blue or immunohistochemical staining. For differentiation into adipocytes, SSC were cultured in DMEM supplemented with 1 x 10<SU-3</SU mu mol/mL dexamethasone, 0.01 mg/mL insulin, 0.5 mu mol/mL IBMX and examined by microscopy after Oil red O staining and reverse transcriptase-polymerase chain reaction (RT-PCR) for gene expression of peroxisome proliferation activation receptor-gamma (PPAR-gamma). Results. After 15 and 21 days of culture in the induction medium for osteoblast differentiation, 75% and 80% chicken SSC differentiated into osteoblasts, as confirmed by Von Kossa's, calcium-cobalt and collagen I antibody staining. After 3 and 7 days of culture in the induction medium for neuron-like cell differentiation, 78% and 85% SSC became neuron-like cells, as confirmed by staining with toluidine blue and the monoclonal antibody against neuron-specific enolase, nestin and glial fibrillary acidic protein. After 7 days of culture in the induction for adipocyte differentiation, 85% SSC differentiated into adipocytes, as confirmed by Oil red O staining and RT-PCT for PPAR-gamma gene expression. Discussion. Our results show that chicken SSC can differentiate into osteoblasts, neuron-like cells and adipocytes under similar conditions as for directional differentiation of mammalian SSC in vitro. The findings show the feasibility of using SSC-derived cells for developmental biology and gene transfer studies in chickens.
引用
收藏
页码:326 / 331
页数:6
相关论文
共 28 条
[1]   TERMINAL NEUROENDOCRINE DIFFERENTIATION OF HUMAN PROSTATE CARCINOMA-CELLS IN RESPONSE TO INCREASED INTRACELLULAR CYCLIC-AMP [J].
BANG, YJ ;
PIRNIA, F ;
FANG, WG ;
KANG, WK ;
SARTOR, O ;
WHITESELL, L ;
HA, MJ ;
TSOKOS, M ;
SHEAHAN, MD ;
NGUYEN, P ;
NIKLINSKI, WT ;
MYERS, CE ;
TREPEL, JB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (12) :5330-5334
[2]   SPERMATOGENESIS FOLLOWING MALE GERM-CELL TRANSPLANTATION [J].
BRINSTER, RL ;
ZIMMERMANN, JW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (24) :11298-11302
[3]   Differentiation of osteoblasts and in vitro bone formation from murine embryonic stem cells [J].
Buttery, LDK ;
Bourne, S ;
Xynos, JD ;
Wood, H ;
Hughes, FJ ;
Hughes, SPF ;
Episkopou, V ;
Polak, JM .
TISSUE ENGINEERING, 2001, 7 (01) :89-99
[4]   Surface modification of poly (D,L-lactic acid) with chitosan and its effects on the culture of osteoblasts in vitro [J].
Cai, KY ;
Yao, KD ;
Cui, YL ;
Lin, SB ;
Yang, ZM ;
Li, XQ ;
Xie, HQ ;
Qing, TW ;
Luo, J .
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, 2002, 60 (03) :398-404
[5]   Human bone cell cultures in biocompatibility testing.: Part II:: effect of ascorbic acid, β-glycerophosphate and dexamethasone on osteoblastic differentiation [J].
Coelho, MJ ;
Fernandes, MH .
BIOMATERIALS, 2000, 21 (11) :1095-1102
[6]   RETRACTED: Generation of pluripotent stem cells from adult human testis (Retracted article. See vol. 512, pg. 338, 2014) [J].
Conrad, Sabine ;
Renninger, Markus ;
Hennenlotter, Joerg ;
Wiesner, Tina ;
Just, Lothar ;
Bonin, Michael ;
Aicher, Wilhelm ;
Buehring, Hans-Joerg ;
Mattheus, Ulrich ;
Mack, Andreas ;
Wagner, Hans-Joachim ;
Minger, Stephen ;
Matzkies, Matthias ;
Reppel, Michael ;
Hescheler, Juergen ;
Sievert, Karl-Dietrich ;
Stenzl, Arnulf ;
Skutella, Thomas .
NATURE, 2008, 456 (7220) :344-U25
[7]   Proliferation and differentiation of spermatogonial stem cells [J].
de Rooij, DG .
REPRODUCTION, 2001, 121 (03) :347-354
[8]   Transplantation of germ cells from rabbits and dogs into mouse testes [J].
Dobrinski, I ;
Avarbock, MR ;
Brinster, RL .
BIOLOGY OF REPRODUCTION, 1999, 61 (05) :1331-1339
[9]   Retinoic acid and development of the retina [J].
Drager, UC ;
McCaffery, P .
PROGRESS IN RETINAL AND EYE RESEARCH, 1997, 16 (03) :323-351
[10]  
ELIZABETH MVJ, 1982, J CELL BIOL, V94, P253