Abstract
Human embryonic stem cells (hESC) have great potential in regenerative medicine, provided that culture systems are established that maintain genomic integrity. Here we describe a comparison of the effects of culture in either physiologic oxygen (2%) or room oxygen (21%) on the hESC lines, H1, H9, and RH1. Physiologic oxygen enabled an average sixfold increase in clone recovery across the hESC lines tested (p <0.001). FACS analysis showed that cells cultured in physiologic oxygen were significantly smaller and less granular. No significant changes had occurred in levels of SSEA4, SSEA1, TRA-1-60, or TRA-1-81. While karyotypic normalcy was maintained in both H1 and H9, the frequency of spontaneous chromosomal aberrations was significantly increased in room oxygen. This increase was not observed in physiologic oxygen. These results clearly demonstrate that physiologic oxygen culture conditions are indispensable for robust hES clone recovery and may enhance the isolation of novel hES lines and transgenic clones.
| Original language | English |
|---|---|
| Pages (from-to) | 16-23 |
| Number of pages | 8 |
| Journal | Cloning and Stem Cells |
| Volume | 8 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2006 |
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ASJC Scopus subject areas
- Biotechnology
- Developmental Biology
- Applied Microbiology and Biotechnology
Cite this
Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities. / Forsyth, Nicholas R.; Musio, Antonio; Vezzoni, Paolo; Simpson, A. Hamish R W; Noble, Brendon S.; McWhir, Jim.
In: Cloning and Stem Cells, Vol. 8, No. 1, 2006, p. 16-23.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities
AU - Forsyth, Nicholas R.
AU - Musio, Antonio
AU - Vezzoni, Paolo
AU - Simpson, A. Hamish R W
AU - Noble, Brendon S.
AU - McWhir, Jim
PY - 2006
Y1 - 2006
N2 - Human embryonic stem cells (hESC) have great potential in regenerative medicine, provided that culture systems are established that maintain genomic integrity. Here we describe a comparison of the effects of culture in either physiologic oxygen (2%) or room oxygen (21%) on the hESC lines, H1, H9, and RH1. Physiologic oxygen enabled an average sixfold increase in clone recovery across the hESC lines tested (p <0.001). FACS analysis showed that cells cultured in physiologic oxygen were significantly smaller and less granular. No significant changes had occurred in levels of SSEA4, SSEA1, TRA-1-60, or TRA-1-81. While karyotypic normalcy was maintained in both H1 and H9, the frequency of spontaneous chromosomal aberrations was significantly increased in room oxygen. This increase was not observed in physiologic oxygen. These results clearly demonstrate that physiologic oxygen culture conditions are indispensable for robust hES clone recovery and may enhance the isolation of novel hES lines and transgenic clones.
AB - Human embryonic stem cells (hESC) have great potential in regenerative medicine, provided that culture systems are established that maintain genomic integrity. Here we describe a comparison of the effects of culture in either physiologic oxygen (2%) or room oxygen (21%) on the hESC lines, H1, H9, and RH1. Physiologic oxygen enabled an average sixfold increase in clone recovery across the hESC lines tested (p <0.001). FACS analysis showed that cells cultured in physiologic oxygen were significantly smaller and less granular. No significant changes had occurred in levels of SSEA4, SSEA1, TRA-1-60, or TRA-1-81. While karyotypic normalcy was maintained in both H1 and H9, the frequency of spontaneous chromosomal aberrations was significantly increased in room oxygen. This increase was not observed in physiologic oxygen. These results clearly demonstrate that physiologic oxygen culture conditions are indispensable for robust hES clone recovery and may enhance the isolation of novel hES lines and transgenic clones.
UR - http://www.scopus.com/inward/record.url?scp=33645766052&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33645766052&partnerID=8YFLogxK
U2 - 10.1089/clo.2006.8.16
DO - 10.1089/clo.2006.8.16
M3 - Article
C2 - 16571074
AN - SCOPUS:33645766052
VL - 8
SP - 16
EP - 23
JO - Cloning and Stem Cells
JF - Cloning and Stem Cells
SN - 1536-2302
IS - 1
ER -