ABSTRACT
Objective: Our knowledge of human neural crest stem cells (NCSCs) is expanding, owing to recent advances in technologies utilizing human-induced pluripotent stem cells (hiPSCs) that generate NCSCs. However, the clinical application of these technologies requires the reduction of xeno-materials. To overcome this significant impediment, this study aimed to devise a novel method to induce NCSCs from hiPSCs without using a feeder cell layer.Materials and Methods: hiPSCs were cultured in feeder-free maintenance media containing the Rho-associated coiled-coil forming kinase inhibitor Y-27632. When the cells reached 50–70% confluence, differentiation was initiated by replacing the medium with knockout serum replacement (KSR) medium containing Noggin and SB431542. The KSR medium was then gradually replaced with increasing concentrations of Neurobasal medium from day 5 to 11.Results: Immunocytochemistry and flow cytometry were performed 12 days after induction of differentiation and revealed that the cells generated from hiPSCs expressed the NCSC markers p75 and HNK-1, but not the hiPSC marker SOX2.Conclusion: These findings demonstrate that hiPSCs were induced to differentiate into NCSCs in the absence of feeder cells.
ABSTRACT
The lack of donor corneal endothelium is a serious impediment to the development of corneal endothelial transplantation,whereas the bioengineered cornea provides an approach to this problem.Functional corneal endothelial cells which differentiated from human embryonic stem cells (ESCs) can,to some extent,relieve the lack of donor corneas,especially for corneal endothelia.At the moment,an optimal way of offering bioengineered-corneal endothelium is to cultivate the corneal endothelial cells in vitro.This is a process of inducing human ESCs to differentiate into neural crest stem cells (NCSCs) and then into corneal endothelial cells in a favorable medium with growth factors and extracellular matrix,which are matched microenvironment of endothelial cells in vitro.However,the inducement condition is still pending and remains for further research.This article reviewed the researching development of bioengineered-corneal endothelium from the effect of microenvironment and the induction of human ESCs.
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Objective To explore the expression of glial cell line-derived neurotrophic factor (GDNF) in rats with spinal cord injury (SCI) after epidermal neural crest stem cells (EPI-NCSCs) transplantation.Method EPI-NCSCs were isolated from GFP transgenic rats for transplantation.The rat SCI model was made by NYU-II impactor (10 g 25 mm) at T10 level.Then 30 SD rats were randomly divided into blank injury group (group A),DMEM transplantation group (group B),and experimental group (group C).The EPI-NCSCs were transplanted into the injured region one week after SCI.In DMEM group,the DMEM/F12 was used to substitute for the EPI-NCSCs.No treatment was done in blank injury group.The locomotor function was appraised by BBB score every week after transplantation.At sixth week after transplantation,GDNF mRNA and protein expression was detected.Result The BBB score in experimental group was significantly higher than the other two groups from two weeks after transplantation (P＜0.05).The expression of GDNF mRNA and protein in experimental group was significantly higher than the other two groups (P＜0.05).There was no significant difference between blank injury group and DMEM transplantation group (P ＞ 0.05).Conclusion The expression of GDNF can be up-regulated by EPI-NCSCs transplantation,which may be one of the mechanisms for EPI-NCSCs repairing SCI.
ABSTRACT
Objective To establish a satisfactory method of isolating,culturing and determining gut neural crest stem cells,which may provide theoretical basis for clinical application.Methods The guts of embryonic mice removed and dissociated were plated into serum-free DMEM/F12 medium.The mitogen-free DMEM/F12 medium supplementd with 10% fetal bovine serum was used to induce differentiation of GNCSCs.Neurospheres and their derivations were determined with immunocytochemical and immunofluorescent staning.Results Neurospheres were generated in the simplified serum-free medium.The staining results showed that enteric neurospheres were GNCSCs and could differentiate into neurons,glial cells and smooth muscle cells by serum-induction.Conclusion GNCSCs have the capacity of self-renewal and proliferation,and give rise to neurons,glial cells and myofibroblasts.