Isolation and characterization of amniotic fluid-derived stem cells in Turner’s syndrome

doi:10.3969/j.issn.2095-4344.2013.14.017

Abstract

Abstract:

BACKGROUND: Great progress has been achieved on studies on isolation, culture and biological characteristics of amniotic fluid-derived mesenchymal stem cells. However, few studies are reported on amniotic fluid-derived mesenchymal stem cells in 45, X/46, XX (Turner’s syndrome).
OBJECTIVE: To develop a simple culture protocol to isolate amniotic fluid-derived mesenchymal stem cells and investigate the biological characteristics of amniotic fluid-derived mesenchymal stem cells.
METHODS: We developed a gradient dilution culture protocol to isolate a population of 45, X/46, XX (Turner’s syndrome) amniotic fluid-derived mesenchymal stem cells from second-trimester amniocentesis. The morphology of amniotic fluid-derived mesenchymal stem cells was observed by light microscope. The karyotype was analyzed. Specific cell surface antigens and cell cycle of the clonal amniotic fluid-derived mesenchymal stem cells at passage 4 were characterized by flow cytometry. Osteogenic differentiation of amniotic fluid-derived mesenchymal stem cells was confirmed by alkaline phosphatase staining and alizarin red staining.
RESULTS AND CONCLUSION: The cultured human amniotic fluid-derived mesenchymal stem cells proliferated rapidly after passage. Karyotype mapping showed abnormal female chromosome type with 45, X/46, XX observed. The amniotic fluid-derived mesenchymal stem cells had an immunophenotype similar to that of common mesenchymal stem cells and were positive for CD29, CD44, CD90 and CD105, but negative for CD34 and CD45. The cell cycle measurement showed that amniotic fluid-derived mesenchymal stem cells cultured in vitro could maintain strong proliferation ability. Alkaline phosphatase staining and alizarin red staining results confirmed that amniotic fluid-derived mesenchymal stem cells could be successfully induced to differentiate into osteocytes under specific culture media. These results demonstrated that amniotic fluid-derived mesenchymal stem cells in 45, X/46, XX (Turner’s syndrome) were successfully isolated and the cells had a great potential of proliferation and showed the characteristics of mesenchymal stem cells.

Key words: stem cells, stem cell culture and differentiation, Turner’s syndrome, amniotic fluid cells, amniotic fluid-derived stem cells, karyotype, osteogenic differentiation, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Gong Yu, Luo Yu-mei, Tian Lin, Liu Hai-bo, Chen Xin-jie, Sun Xiao-fang, Chen Yao-yong. Isolation and characterization of amniotic fluid-derived stem cells in Turner’s syndrome[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(14): 2586-2591.

Figures/Tables 5

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