Bone morphogenetic protein 4 promotes craniofacial neural crest induction from human pluripotent stem cells

Abstract

Neural crest (NC) cells are a group of cells located in the neural folds at the boundary between the neural and epidermal ectoderm. Cranial NC cells migrate to the branchial arches and give rise to the majority of the craniofacial region, whereas trunk and tail NC cells contribute to the heart, enteric ganglia of the gut, melanocytes, sympathetic ganglia, and adrenal chromaffin cells. Positional information is indispensable for the regulation of cranial or trunk and tail NC cells. However, the mechanisms underlying the regulation of positional information during human NC induction have yet to be fully elucidated. In the present study, supplementation of bone morphogenetic protein (BMP) 4 in defined serum-free culture conditions including fibroblast growth factor-2 and Wnt3a from day 8 after NC specification induced the expression of cranial NC markers, AP2a, MSX1, and DLX1, during NC cell differentiation from human pluripotent stem cells. On the other hand, the proportion of cells expressing p75 NTR or HNK1 decreased compared with that of cells cultured without BMP4, whereas gene expression analysis demonstrated that the expression levels of cranial NC-associated genes increased in BMP4-treated NC cells. These BMP4-treated NC cells were capable of differentiation into osteocytes and chondrocytes. The results of the present study indicate that BMP4 regulates cranial positioning during NC development.

Establishment and neural differentiation of neural crest-derived stem cells from human dental pulp in serum-free conditions

“…35 However, the observed hDPC-BMP4 NCSC phenotype, as well as the improved changes observed during their neural differentiation in vitro, support the important role of BMP4 in sensory neuron and cranio-facial neural crest induction. 17,30,42,43 To provide further evidence of the importance of BMP4 for the NCSC phenotype and neural differentiation, we also tested the effect of supplementing hDPC- Notably, the characterization of dental-related human NCSCs, including those derived from gingival tissue, periodontal tissue, exfoliated deciduous teeth, and adult human dental pulp, has gained increasing interest for their potential use in neurodegenerative disorders. Nevertheless, most reports are limited to describing the molecular resemblance to neural crest cells, while their actual differentiation capacity is usually left untested, masking their actual translational potential for regenerative medicine.…”

In vitro models of cranial neural crest development toward toxicity tests: frog, mouse, and human

“…In our recent study (Mimura et al, 2016), we demonstrated a two-step NC induction method: (1) the activation of WNT and FGF signaling on days 0-8 for early NC specification and (2) the activation of BMP signaling in combination with WNT and FGF signaling on days 8-12 ( Figure 3a). Activating BMP signaling at specific time points appears critical for the induction of cranial mesenchyme from early NC cells (Figure 3b).…”

“…Microarray analysis demonstrated the upregulation of cranial NC genes in the induced craniofacial mesenchyme (Figure c). These BMP‐treated NCCs can differentiate into osteocytes and chondrocytes (Mimura et al , ). These results indicate that BMP signaling regulates cranial positioning during NC differentiation from hPSCs.…”

“…Blue, red, and green lines indicate early NC , cranial NC , and migratory and posterior NC marker genes, respectively. Source : Panels ( b – c ) modified from Mimura et al , .…”

Contribution of cranial neural crest cells to mouse skull development

“…in a 3D bioengineering microenvironment where neural crest cells are encapsulated in gelatin-based photo-cross-linkable hydrogels and cultured for about 3 weeks to efficiently drive differentiation (Namkoong et al, 2016). In addition, CNCs supplemented with BMP4 in the culture media are capable of differentiating into osteocytes and chondrocytes (Mimura et al, 2016). Mouse embryonic stem cells can be selectively differentiated into cranial NC stem cells, which can further differentiate into other cell types including osteoblasts (Minamino et al, 2015).…”