Human adipose derived stem cells (hASCs) can be easily isolated and their plasticity has been well characterized. Several TGF-β superfamily ligands can direct hASCs towards chondrocytes. However, these ligands are difficult to purify and expensive. We have developed a library of Activin/BMP2 chimeric ligands (AB2 ligands) by systematically mixing their sequence segments and have tested their chondrogenic potential in hASCs. Cells cultured in monolayer or in a pellet culture system were incubated with a chemically defined medium supplemented with the chimeric ligands for 4 or 6. weeks and showed higher expression levels of type II collagen, aggrecan, and Sox9 mRNAs when compared with control and non-treated cells. Moreover, toluidine blue, alcian blue, and Masson's trichrome staining was markedly increased in treated cells, both in cell pellet and monolayer assays. In addition, immunohistochemical staining for detection of type I collagen, type II collagen, and Sox 9 demonstrated the acquisition of a chondrogenic phenotype in both culture systems. We present here an inexpensive and robust protocol for differentiation of hASCs towards chondrocytes in a reproducible and highly efficient manner. The AB2 ligands employed are easily produced and have properties that may become useful in cell therapy.
|Original language||English (US)|
|Number of pages||13|
|Journal||Stem Cell Research|
|State||Published - May 2013|
Bibliographical noteFunding Information:
Macarena Peran was supported first by the Spanish Ministry of Education through the National Human Resources Mobility National Plan I-D + i 2008–2011 and continued with a grant from Jaen University, Spain . J.A.M.: Junta de Andalucía, excellence project number CTS-6568. Work in the laboratory of J.C.I.B. was supported by grants from TERCEL-ISCIII-MINECO, Fundacion Cellex, CIBER-BBN , The Leona M. and Harry B. Helmsley Charitable Trust, G. Harold and Leila Y. Mathers Charitable Foundation and The Ellison Medical Foundation.
ASJC Scopus subject areas
- Developmental Biology
- Cell Biology