Transplantation of Neural Crest-Like Cells Derived from Induced Pluripotent Stem Cells Improves Diabetic Polyneuropathy in Mice

Abstract

Impaired vascularity and nerve degeneration are the most important pathophysiological abnormalities of diabetic polyneuropathy (DPN). Therefore, regeneration of both the vascular and nervous systems is required for the treatment of DPN. The neural crest (NC) is a transient embryonic structure in vertebrates that differentiates into a vast range of cells, including peripheral neurons, Schwann cells, and vascular smooth muscle cells. In this study, we investigated the ability of transplantation of NC-like (NCL) cells derived from aged mouse induced pluripotent stem (iPS) cells in the treatment of DPN. iPS cells were induced to differentiate into neural cells by stromal cell-derived inducing activity (SDIA) and subsequently supplemented with bone morphogenetic protein 4 to promote differentiation of NC lineage. After the induction, p75 neurotrophin receptor-positive NCL cells were purified using magnetic-activated cell sorting. Sorted NCL cells differentiated to peripheral neurons, glial cells, and smooth muscle cells by additional SDIA. NCL cells were transplanted into hind limb skeletal muscles of 16-week streptozotocin-diabetic mice. Nerve conduction velocity, current perception threshold, intraepidermal nerve fiber density, sensitivity to thermal stimuli, sciatic nerve blood flow, plantar skin blood flow, and capillary number-to-muscle fiber ratio were evaluated. Four weeks after transplantation, the engrafted cells produced growth factors: nerve growth factor, neurotrophin 3, vascular endothelial growth factor, and basic fibroblast growth factor. It was also confirmed that some engrafted cells differentiated into vascular smooth muscle cells or Schwann cell-like cells at each intrinsic site. The transplantation improved the impaired nerve and vascular functions. These results suggest that transplantation of NCL cells derived from iPS cells could have therapeutic effects on DPN through paracrine actions of growth factors and differentiation into Schwann cell-like cells and vascular smooth muscle cells.

Insights into neural crest development and evolution from genomic analysis

“…There is also the promise that induced neural crest cells could be used for cell therapy for diseases that involve failure or impairment of neural crest-derived tissue Barraud et al 2010). For instance, transplantation of induced ''neural crest-like cells'' to diabetic mice resulted in improvement of the impaired nerve and vascular functions that result from diabetic neuropathy (Okawa et al 2012). One would imagine that engineered neural crest cells could be used in such fashion to treat neurodegenerative disorders that affect peripheral nervous system, among others.…”

Pluripotent Stem Cells for Schwann Cell Engineering

“…NCSCs were seeded in nerve conduits, implanted in a rat sciatic nerve injury model, and after 1 month formed S100β-expressing cells indicating in vivo glial differentiation. Okawa et al used PA6 (SDIA) and BMP4 to differentiate GFP-positive mouse iPSCs towards NC-like cells, which were then FACS-sorted for p75 NTR expression and implanted intramuscularly in a mouse model for diabetic neuropathy [124]. Interestingly, 4 weeks after transplantation, GFP-positive S100β-expressing SC-like cells could be detected, indicating intramuscular glial differentiation.…”

“…Addition of SHH and FGF8 to differentiating pluripotent cells can be used to induce neural rosette formation [118], but is often avoided as it can suppress development of dorsal neural tissues and promote ventralization [122]. Some authors use BMP to induce NC formation [123,124]. Although early BMP antagonism (e.g.…”

Is There a Role for Neural Crest Stem Cells in Periodontal Regeneration?

“…Induced pluripotent stem cells (iPSC) are amongst the most promising cell sources for tissue engineering because iPSC show high multi-differentiation and self-renewal capabilities [70,71]. Recently, several studies have reported the successful generation of NCSC from iPSC [72][73][74][75][76]. Interestingly, iPSC-derived NCSC showed no tumor formation after their transplantation into nude mice and could differentiated into odontoblast-like cells that expressed odontoblast markers, DSP, Pax9, Msx1, and Lhx6 [76].…”