Abstract
Introduction The adult neural crest-derived stem cells (NCSCs) have significant perspectives for use in regenerative medicine. The most attractive sources for adult NCSC isolation are the hair follicles (HF) and skin dermis (SD) because of easy access and minimally invasive biopsy. The aim of this study was to compare the biological properties of HF- and SD-derived NCSCs after their large-scale expansion. Methods The conventional explant method was used to obtain HF NCSCs. For the isolation of SD NCSCs, a new combined technique consisting of preplating and subsequent culturing in 3D blood plasma-derived fibrin hydrogel was applied. The studied cells were characterized by flow cytometry, ICC, qPCR, Bio-Plex multiplex assay, and directed multilineage differentiation assays. Results We have obtained both adult SD and HF NCSCs from each skin sample (n = 5). Adult SD and HF NCSCs were positive for key neural crest markers: SOX10, P75 (CD271), NESTIN, SOX2, and CD349. SD NCSCs showed a higher growth rate during the large-scale expansion compared to HF NCSCs (p < 0.01). Final population of SD NCSCs also contained more clonogenic cells (p < 0.01) and SOX10+, CD271+, CD105+, CD140a+, CD146+, CD349+ cells (p < 0.01). Both HF and SD NCSCs had similar gene expression profiling and produced growth factors, but some quantitative differences were detected. Adult HF and SD NCSCs were able to undergo directed differentiation into neurons, Schwann cells, adipocytes, and osteoblasts. Conclusion The HF and SD are suitable sources for large-scale manufacturing of adult NCSCs with similar biological properties. We demonstrated that the NCSC population from SD was homogenous and displayed significantly higher growth rate than HF NCSCs. Moreover, SD NCSC isolation is cheaper, easier, and minimally time-consuming method.
Highlights
The adult neural crest-derived stem cells (NCSCs) have significant perspectives for use in regenerative medicine
Specific growth medium provides suitable conditions for the preferable growth of the target NCSC population comparing to keratinocytes and melanocytes, which can be obtained from the same hair follicles (HF) explant
The homogenous NCSC population is obtained by differential trypsinization when passaging the primary cell culture, as described earlier [45]
Summary
The adult neural crest-derived stem cells (NCSCs) have significant perspectives for use in regenerative medicine. Adult HF and SD NCSCs were able to undergo directed differentiation into neurons, Schwann cells, adipocytes, and osteoblasts. The Canadian scientist Brain Hall assumed that NC is a fourth embryonic layer taking into consideration its role in Vertebrates ontogenesis and phylogenesis [3] This concept is becoming increasingly common in the scientific community. There are several domains within NC, among which the cells of the cranial neural crest possess the most wide-ranging potential for multilineage differentiation. They give rise to ectomesenchyme (i.e., different mesenchymal cell types, like adipocytes, osteoblasts, and chondrocytes), melanocytes, neurons, and glia of the peripheral nervous system [4]
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