Nature Biotechnology
20, 933 - 936 (2002)
Published online: 5 August 2002; | doi:10.1038/nbt726
Human feeders support prolonged undifferentiated growth of human inner
cell masses and embryonic stem cellsMark Richards1, Chui-Yee Fong1, Woon-Khiong Chan2, Peng-Cheang Wong1
& Ariff Bongso11
Department of Obstetrics and Gynaecology, National
University of Singapore, Kent Ridge, Singapore
119260. 2
Department of Biological Sciences, National
University of Singapore, Kent Ridge, Singapore
119260.
Correspondence should be addressed to Ariff Bongso obgbongs@nus.edu.sgPrevious reports have demonstrated the growth of undifferentiated
human embryonic stem (HES) cells on mouse embryonic fibroblast (MEF) feeders
and on laminin- or Matrigel-coated plastic surfaces supplemented with
MEF-conditioned medium1,
2,
3. These xenosupport systems run the
risk of cross-transfer of animal pathogens from the animal feeder, matrix, or
conditioned medium to the HES cells, thus compromising later clinical
application. Here we show that human fetal and adult fibroblast feeders support
prolonged undifferentiated HES cell growth of existing cell lines and are
superior to cell-free matrices (collagen I, human extracellular matrix,
Matrigel, and laminin) supplemented with human or MEF feeder−conditioned
medium. Additionally, we report the derivation and establishment of a new HES
cell line in completely animal-free conditions. Like HES cells cultured on MEF
feeders, the HES cells grown on human feeders had normal karyotypes, tested
positive for alkaline phosphatase activity, expressed Oct-4 and cell surface
markers including SSEA-3, SSEA-4, Tra 1-60, and GCTM-2, formed teratomas in
severely combined immunodeficient (SCID) mice, and retained all key
morphological characteristics. Human feeder−supported HES cells should
provide a safer alternative to existing HES cell lines in therapeutic
applications.
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