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
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Mentions: Generation of osteochondrogenic ectomesenchyme from hPSCs first requires specification of cranial neural crest-like progeny. In order to reproducibly generate strong chondrogenic activity from hPSCs, we first optimized the method for specification of neural crest using CD271 as the readout. Fluorescence-activated cell sorting (FACS) analyses revealed that when H9 hESCs and BJ5 hiPSCs were differentiated in CDM using the conventional embryoid body (EB)-forming culture or 2D differentiation culture in the presence of SB431542 or other Nodal/Activin/TGFβ signaling inhibitor (e.g., A83-01 or SJN2511), they generated progeny that expressed CD271 (Figures 1A and S1). The CD271hi cell population, but not the CD271lo/− progeny, also expressed PDGFRα at low levels. Both populations lacked CD73 and CD13. The effect of SB431542 was dose dependent at least up to 10 μM (Figure S1D).
Affiliation: Institute of Molecular Medicine, The University of Texas Health Science Center at Houston Medical School, Houston, TX 77030, USA.