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Embryonic Stem Cells |
1 Cellartis AB, Göteborg, Sweden; Stem Cell Center, Lund University, Lund, Sweden
2 Cellartis AB, Göteborg, Sweden
3 Reproductive Medicine, Department of Obstetrics and Gynaecology, Institute for Health of Women and Children, Sahlgrenska Academy, Göteborg, Sweden
4 Stem Cell Center, Lund University, Lund, Sweden
* To whom correspondence should be addressed. E-mail: henrik.semb{at}med.lu.se.
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Abstract |
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Elimination of all animal material during both the derivation and long-term culture of hESC is necessary prior to future application of human embryonic stem cells (hESC) in clinical cell therapy. The potential consequences of transplanting xenocontaminated hESC into patients, such as an increased risk of graft rejection [1] and the potential transfer of non-human pathogens, make existing hESC lines unsuitable for clinical applications. To avoid xeno-contamination during derivation and culture of hESC, we first developed a xeno-free medium supplemented with human serum (HS), which supports long-term (>50 passages) culture of hESC in an undifferentiated state. To enable derivation of new xeno-free hESC, we also established xeno-free human foreskin fibroblast (HFF) feeders and replaced immunosurgery, which involves the use of guinea pig complement, with a modified animal-product-free derivation procedure. Here we report the establishment and characterization (>20 passages) of a xeno-free pluripotent diploid normal hESC line, SA611.
Key Words. human embryonic stem cell, human serum, human feeders, clinical therapies
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