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Original Article |
1 Centre for Stem Cell Biology and Developmental Genetics, Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne, UK
2 School of Biological and Biomedical Sciences, University of Durham, Durham, UK
3 Centre for Stem Cell Biology and Developmental Genetics, Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne, UK; School of Biological and Biomedical Sciences, University of Durham, Durham, UK
4 Institute of Human Genetics, University of Newcastle, Newcastle upon Tyne, UK
* To whom correspondence should be addressed. E-mail: miodrag.stojkovic{at}ncl.ac.uk.
| Abstract |
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One of the most frequently used matrix for feeder-free growth of undifferentiated hESC is Matrigel which supports attachment and growth of undifferentiated hESC in the presence of MEF-conditioned medium. Unfortunately, application of Matrigel or medium conditioned by mouse embryonic feeder cells is not ideal for potential medical application of hESC because xenogeneic pathogens can be transmitted through culture conditions. We demonstrate here that human serum (HS) as matrix and medium conditioned by differentiated hESC (hES-dF) reduce exposure of hESC to animal ingredients and provides a safer direction towards completely animal-free conditions for application, handling and understanding of hESC biology. At the same time, hESC cells grown under these conditions maintain all hESC features after prolonged culture, including the developmental potential to differentiate into representative tissues of all three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype.
Key Words. human embryonic stem cells, pluripotency, differentiation, feeder-free
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