Long-term expandable SOX9+ chondrogenic ectomesenchymal cells from human pluripotent stem cells.
Bottom Line: When "primed" with TGFβ, such cells efficiently formed translucent cartilage particles, which were completely mineralized in 12 weeks in immunocompromized mice.Ectomesenchyme is a source of many craniofacial bone and cartilage structures.The method we describe for obtaining a large quantity of human ectomesenchymal cells will help to model craniofacial disorders in vitro and potentially provide cells for the repair of craniofacial damage.
Affiliation: Institute of Molecular Medicine, The University of Texas Health Science Center at Houston Medical School, Houston, TX 77030, USA.Show MeSH
Related in: MedlinePlus
Mentions: To determine the cellular origin of the CD271+CD73+ ectomesenchymal cells expanded under FGF2+SB431542 conditions, the CD271hiCD73− and CD271loCD73− progeny, developed from H9 hESCs and BJ5 hiPSCs, were sorted by FACS (Figures 1C and S1H) and then cultured in CDM in the presence of FGF2 and one of three Nodal/Activin/TGFβ inhibitors, SB431542, A83-01, or SJN2511. While FGF2 alone supported the accumulation of CD271−CD73+ cells from either progeny, the FGF2+SB431542, FGF2+A83-01, and FGF2+SJN2511 conditions preferentially supported the development of CD271+CD73+ cells from the sorted CD271hiCD73− neural crest-like progeny (Figures 3B and S4G). Concentrations of SB431542 between 1 and 3 μM were sufficient for the stable maintenance of CD271+CD73+ cells (Figure S3C). On the other hand, most of the CD271loCD73− progeny did not survive, even under FGF2+SB431542 conditions, and the derived mesenchymal cells were a mixture of CD271+CD73+ and CD271−CD73+ cells (Figures 3B and S4G) from which the CD271+CD73+ cells were later lost.
Affiliation: Institute of Molecular Medicine, The University of Texas Health Science Center at Houston Medical School, Houston, TX 77030, USA.