Abstract
The majority of human embryonic stem cell (hESC) lines have been derived and grown using mouse or human feeder cells, or using Matrigel®, an animal derivative rich in extracellular matrix (ECM) proteins. However, reliance on feeder layers and animal products limits the manipulation and clinical application of hESC. Alternatively, human fibroblasts produce an ECM which could be employed to coated plates and be easily sterilized. We have shown that hESC grown on this matrix and in the presence of medium conditioned by fibroblast cells maintain markers of pluripotency, including expression of cell surface proteins (SSEA3, SSEA4, TRA-1-60, TRA-1-81), alkaline phosphatase activity, and specific intracellular markers (NANOG, OCT, REX1). Moreover, hESC cultured on this novel human-derived ECM display a normal karyotype. This growth system reduces exposure of hESC to feeder layers and animal ingredients, thereby limiting the risk of pathogenic contamination and additionally facilitating manipulation of hESCs.
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Acknowledgments
The authors would like to thank Dr. Deborah Burks and Tugce Pehlivan for critical reading of the manuscript and Dario Melguizo, Sonia Prado, Dr. Angel Ayuso-Sacido, Dr. X. Chen Xiong, and Petra Stojkovic for technical support. This work was supported by funds for research in the field of Regenerative Medicine from the Regional Government Health Department (Generalitat Valenciana), the Instituto Carlos III belonging to the Spanish Ministry of Health and Consumer Affairs.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC 2006
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Escobedo-Lucea, C., Stojkovic, M. (2009). Growth of Human Embryonic Stem Cells Using Derivates of Human Fibroblasts. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 584. Humana Press. https://doi.org/10.1007/978-1-60761-369-5_3
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DOI: https://doi.org/10.1007/978-1-60761-369-5_3
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