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JBMR Plus (WOA)
24734039, 2023, 5, Downloaded from https://asbmr.onlinelibrary.wiley.com/doi/10.1002/jbm4.10737 by Cochrane Japan, Wiley Online Library on [15/05/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
to the formation of growth plate structures. These observations
might support previous reports describing stem cell populations
in proliferating chondrocytes.(43,44) Although the difference was
statistically significant only in the case of 414C2, the proportion
of the bone area in each tissue was larger in COL X-deficient tissues than in parental tissues, suggesting the acceleration of differentiation. The ectopic expression of SOX9 in hypertrophic
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Cells at day 28 are already positive for IHH, a marker for prehypertrophic chondrocytes and also for RUNX2. No proliferating
chondrocytes were observed in collected pellets, suggesting
that most cells at day 28 in vitro may lose the capacity to proliferate and only the differentiation process continued in vivo.
The presence of HNA-positive osteoblasts and osteocytes in
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with osteogenic potentials existed in day 28 pellets or that
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There are several limitations in our hiPSC differentiation system. Most importantly, it lacks the well-organized regulatory network in growth plates such as the IHH-PTHrP pathway,(1) which
may result in a nonphysiological differentiation process.
Mechanical factors, which are another important factor for the
metabolic regulation of the growth plate,(2) are also missing. Further modifications of our system are required to faithfully recapitulate the physiological process of chondrocyte differentiation.
In conclusion, COL X deficiency is dispensable for columnar
differentiation processes including hypertrophic differentiation
and the endochondral ossification of chondrocytes induced
from hiPSCs. COL X deficiency may have, however, an effect on
facilitating the differentiation process through an unknown
mechanism. The combination of gene-edited human iPSCs and
in vitro and in vivo assay systems for hypertrophic differentiation
and growth plate-like structures is useful for investigating the
role of molecules involved in the differentiation and also the
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