title: Activin/BMP2 chimeric ligands direct adipose-derived stem cells to chondrogenic differentiation.
abstract: 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.
authors.affiliation: Gene Expression Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
authors.forename: Macarena
authors.initials: M
authors.lastname: Peran
authors.forename: Sergio
authors.initials: S
authors.lastname: Ruiz
authors.forename: Witek
authors.initials: W
authors.lastname: Kwiatkowski
authors.forename: Juan Antonio
authors.initials: JA
authors.lastname: Marchal
authors.forename: Sheng-Lian
authors.initials: SL
authors.lastname: Yang
authors.forename: Antonia
authors.initials: A
authors.lastname: Aranega
authors.forename: Senyon
authors.initials: S
authors.lastname: Choe
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Izpisua Belmonte
mesh_terms: D028341:Activins / Q000235:genetics / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology*; D000328:Adult; D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D020219:Chondrogenesis / Q000187:drug effects; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D008024:Ligands; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D053674:Aggrecans; D055396:Bone Morphogenetic Protein 2; D024043:Collagen Type II; D008024:Ligands; D011993:Recombinant Fusion Proteins; D055758:SOX9 Transcription Factor; D028341:Activins