| | |  | | | | Record from Web of Science® | | | | | | |
| Feeder-independent culture of human embryonic stem cells |
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| Author(s): Ludwig TE (Ludwig, Tenneille E.), Bergendahl V (Bergendahl, Veit), Levenstein ME (Levenstein, Mark E.), Yu JY (Yu, Junying), Probasco MD (Probasco, Mitchell D.), Thomson JA (Thomson, James A.) |
| Source: NATURE METHODS Volume: 3 Issue: 8 Pages: 637-646 Published: AUG 2006 |
| Times Cited: 58 References: 17 |
| Abstract: We recently reported the development of TeSR1, a serum-free, animal product-free medium that supports the derivation and long-term feeder-independent culture of human embryonic stem cells(1). Although the derivation of new human embryonic stem cell lines in those defined conditions offered an important proof of principle, the costs of some of the defined components in that culture system made it impractical for everyday research use. Here we describe modifications to the medium (mTeSR1) that include the use of animal-sourced proteins (bovine serum albumin (BSA) and Matrigel) and cloned zebrafish basic fibroblast growth factor (zbFGF). We include a simple protocol that allows purification of up to 100 mg zbFGF in less than three days (Fig. 1), an amount sufficient to make 1,000 I of mTeSR1 medium. The modifications presented here make mTeSR1 practical for routine research use, and the protocols presented are those currently used in our laboratory for standard human embryonic stem cell culture. |
| Document Type: Article |
| Language: English |
| Reprint Address: Ludwig, TE (reprint author), Univ Wisconsin, Genome Ctr Wisconsin, 425 Henry Mall, Madison, WI 53706 USA |
Addresses:
1. Univ Wisconsin, Genome Ctr Wisconsin, Madison, WI 53706 USA
2. WiCell Res Inst, Madison, WI 53707 USA
3. Univ Wisconsin, Grad Sch, Natl Primate Res Ctr, Madison, WI 53705 USA
4. Univ Wisconsin, Sch Med, Dept Anat, Madison, WI 53706 USA |
| Publisher: NATURE PUBLISHING GROUP, MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND |
| Subject Category: Biochemical Research Methods |
| IDS Number: 071JT |
| ISSN: 1548-7091 |
| DOI: 10.1038/NMETH902 |
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