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Growth and identification of human amniotic fluid stem cells and analysis of their influencing factors

Published online by Cambridge University Press:  24 April 2009

Wang Han
Affiliation:
College of Veterinary Medicine, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Shaanxi Stem Cell Engineering and Technology Research Center, Northwest Agriculture and Forestry University, Yangling 712100, China The College of Life Science and Chemistry, Xinjiang Normal University, Wulumuqi 830054, China
Dou Zhong-Ying
Affiliation:
College of Veterinary Medicine, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Shaanxi Stem Cell Engineering and Technology Research Center, Northwest Agriculture and Forestry University, Yangling 712100, China
Wang Hua-Yan*
Affiliation:
College of Veterinary Medicine, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Shaanxi Stem Cell Engineering and Technology Research Center, Northwest Agriculture and Forestry University, Yangling 712100, China
*
*Corresponding author. E-mail: [email protected]

Abstract

Amniotic fluid stem (AFS) cells isolated from human amniotic fluid were cultured and the factors affecting the primary culture of these cells were investigated. The isolated AFS cells express both embryonic stem cell markers, such as Oct-4, and adult stem cell markers, such as CD29, and possess a high proliferation capability. Moreover, isolated AFS cells were able to differentiate into Nestin-positive and α-actin-positive cells. There were several factors, including the date of gestation, the level of blood contamination and the volume of amniotic fluid, which could significantly affect the attachment time and numbers of AFS cells in primary culture. The results showed that the cell attachment time (4.7±0.6 days) during the second trimester of gestation was significantly different from that during the third trimester of gestation (6±0.5 days) (P<0.05), suggesting that cells collected from the fluid during the third trimester of gestation needed a longer attachment time. Blood contamination could significantly affect the cell attachment time. The attachment time of the brown-coloured fluid group (10.8±0.3 days) was significantly different from the blood-cell-free fluid group (6±0.5 days) and the blood-cell group (6.3±0.6 days) (P<0.05). The volume of amniotic fluid influenced, to some extent, both cell attachment numbers and time. With the increase in amniotic fluid volume, cell attachment numbers significantly increased (P<0.05), and cell attachment time extended, but not significantly (P>0.05). The present studies systematically examined the factors that influence the primary culture of human AFS cells and provided useful data on AFS cell research. Additionally, the isolated AFS cells maintain the capacity for differentiation into other cell types and are able to become seed cells for clinical applications.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2009

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References

Chang, HC and Jones, OW (1991) Enhancement of amniocyte growth on a precoated surface. Prenatal Diagnosis 11(6): 357370.Google Scholar
Chiavegato, A, Bollini, S, Pozzobon, M, et al. (2007) Human amniotic fluid-derived stem cells are rejected after transplantation in the myocardium of normal, ischemic, immuno-suppressed or immuno-deficient rats. Journal of Molecular and Cellular Cardiology 42(4): 746759.CrossRefGoogle ScholarPubMed
De Coppi, P, Bartsch, G, Siddiqui, MM, et al. (2007) Isolation of amniotic stem cell lines with potential for therapy. Nature Biotechnology 25(1): 100106.CrossRefGoogle ScholarPubMed
Kim, J, Lee, Y, Kim, H, et al. (2007) Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells. Cell Proliferation 40(1): 7590.CrossRefGoogle ScholarPubMed
Malaren, A (2001) Ethical and social considerations of stem cell research. Nature 414(6859): 129131.Google Scholar
Meng, GL, Shang, KG and Ding, MX (2002) Current state of establishing and maintaining human embryonic stem cell lines and key problems in these studies. Chinese Journal of Biotechnology 18(2): 131135.Google ScholarPubMed
Passier, R and Mummery, C (2003) Origin and use of embryonic and adult stem cells in differentiation and tissue repair. Cardiovascular Research 58(2): 324335.CrossRefGoogle ScholarPubMed
Prusa, AR and Hengstschlager, M (2002) Amniotic fluid cells and human stem cell research – a new connection. Medical Science Monitor 8(11): 253257.Google ScholarPubMed
Rideout, WM, Hochedlinger, K, Kyba, M, Daley, GQ and Jaenisch, R (2002) Correction of a genetic defect by nuclear transplantation and combined cell and gene therapy. Cell 109(1): 1727.CrossRefGoogle ScholarPubMed
Seguin, LR and Palmer, CG (1983) Variables influencing growth and morphology of colonies of cells from human amniotic fluid. Prenatal Diagnosis 3(2): 107116.CrossRefGoogle ScholarPubMed
Sikkema-Raddatz, B, van Echten, J, van der Vlag, J, Buys, C and te Meerman, GJ (2002) Minimal volume of amniotic fluid for reliable prenatal cytogenetic diagnosis. Prenatal Diagnosis 22(2): 164165.Google Scholar
Tsai, MS, Hwang, SM, Tsai, YL, Cheng, FC, Lee, JL and Chang, YJ (2006) Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells. Biology of Reproduction 74(3): 545551.CrossRefGoogle ScholarPubMed
Zwaka, TP and Thomson, JA (2003) Homologous recombination in human embryonic stem cells. Nature Biotechnology 21(3): 319324.CrossRefGoogle ScholarPubMed