A team of researchers at the University of Wisconsin-Madison reports the development of a fully defined embryonic or pluripotent stem cells culture system.
[R&D Systems has an excellent defined media, StemXVivo culture matrix, composed of a mixture of recombinant human adhesion molecules for the culture of stem/progenitor cells.]Writing this week (Nov. 14, 2010) in
Nature Methods, a team led by Laura Kiessling, unveiled an inexpensive system that takes much of the guess work out of culturing the all-purpose cells.
"It's a technology that anyone can use," says Kiessling. "It's very simple."
The new culture system utilizes a synthetic, chemically made substrate of protein fragments, peptides, which have an affinity for binding with stem cells. Used in combination with a defined growth medium, the system devised by the Wisconsin team can culture cells in their undifferentiated states for up to three months and possibly longer. The system, according to the new report, also works for induced pluripotent stem cells, the adult cells genetically reprogrammed to behave like embryonic stem cells.
Figure 3 (Lim et al.): Synthetic surfaces support the long-term culture of pluripotent stem cells.
(a) Immunostaining of H9 hES cells cultured for three months in mTeSR with ROCK inhibitor on the synthetic surface for Oct-4 and SSEA-4, and stained with DAPI. Scale bar, 200 μm. (b,c) Immunostaining of DF19-9 7T iPS cells cultured for 2.5 months in mTeSR with ROCK inhibitor on the synthetic surface for nanog and E-cadherin (b) or for Sox2 and Tra-1-60 R&D Systems reagents used in this study:
recombinant proteins:
Activin ABMP-4NogginPlates:
Vitronectin-coated plates
Antibodies:
Oct-4NanogSox-2β-III tubulinFoxA2Fatty acid binding protein 4Alkaline phosphatase, APC-conjugated
SSEA-1, PE–conjugated
Joseph R Klim, Lingyin Li, Paul J Wrighton, Marian S Piekarczyk & Laura L Kiessling. A defined glycosaminoglycan-binding substratum for human pluripotent stem cells.
Nature Methods, 14 November 2010 DOI: 10.1038/nmeth.1532[News adapted from this UW-Madison press release]
Szabo et al. (7 Nov 2010; doi:10.1038/nature09591) transduced human skin fibroblasts with the pluripotency factor OCT4 and grew them with different cytokines to convert them into hematopoietic progenitors and mature blood cells. After a few days, the cells expressed the pan- haematopoietic factor CD45 and eventually gave rise to all blood cell types (myeloid, erythroid and megakaryocytic lineages), depending on the cytokines added to the medium.
