अलगाव और तंत्रिका शिखा मानव बालों के रोम से स्टेम सेल के संस्कृति
Summary
इस अनुच्छेद के अलगाव और मानव बालों के रोम से तंत्रिका शिखा स्टेम कोशिकाओं की संस्कृति के लिए एक मजबूत प्रोटोकॉल प्रस्तुत करता है.
Abstract
Hair follicles undergo lifelong growth and hair cycle is a well-controlled process involving stem cell proliferation and quiescence. Hair bulge is a well-characterized niche for adult stem cells1. This segment of the outer root sheath contains a number of different types of stem cells, including epithelial stem cells2, melanocyte stem cells3 and neural crest like stem cells4-7. Hair follicles represent an accessible and rich source for different types of human stem cells. We and others have isolated neural crest stem cells (NCSCs) from human fetal and adult hair follicles4,5. These human stem cells are label-retaining cells and are capable of self-renewal through asymmetric cell division in vitro. They express immature neural crest cell markers but not differentiation markers. Our expression profiling study showed that they share a similar gene expression pattern with murine skin immature neural crest cells. They exhibit clonal multipotency that can give rise to myogenic, melanocytic, and neuronal cell lineages after in vitro clonal single cell culture. Differentiated cells not only acquire lineage-specific markers but also demonstrate appropriate functions in ex vivo conditions. In addition, these NCSCs show differentiation potential toward mesenchymal lineages. Differentiated neuronal cells can persist in mouse brain and retain neuronal differentiation markers. It has been shown that hair follicle derived NCSCs can help nerve regrowth, and they improve motor function in mice transplanted with these stem cells following transecting spinal cord injury8. Furthermore, peripheral nerves have been repaired with stem cell grafts9, and implantation of skin-derived precursor cells adjacent to crushed sciatic nerves has resulted in remyelination10. Therefore, the hair follicle/skin derived NCSCs have already shown promising results for regenerative therapy in preclinical models.
Somatic cell reprogramming to induced pluripotent stem (iPS) cells has shown enormous potential for regenerative medicine. However, there are still many issues with iPS cells, particularly the long term effect of oncogene/virus integration and potential tumorigenicity of pluripotent stem cells have not been adequately addressed. There are still many hurdles to be overcome before iPS cells can be used for regenerative medicine. Whereas the adult stem cells are known to be safe and they have been used clinically for many years, such as bone marrow transplant. Many patients have already benefited from the treatment. Autologous adult stem cells are still preferred cells for transplantation. Therefore, the readily accessible and expandable adult stem cells in human skin/hair follicles are a valuable source for regenerative medicine.
Protocol
Discussion
सेल अलगाव और संस्कृति वर्णित विधियों प्रतिलिपि प्रस्तुत करने योग्य और मजबूत कर रहे हैं. हम एक व्यापक आयु सीमा के पार व्यक्तियों के दर्जनों से NCSCs उत्पन्न किया है. हालांकि यह सबसे अच्छा है ऊतक फसल के बाद ?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
यह काम NIH अनुदान R01AR054593 और Xu के R01AR054593-S1 द्वारा समर्थित है.
Materials
| Name of the reagent | Company | Catalogue number |
| DMEM | Invitrogen | 11965-092 |
| DMEM/F12 | Invitrogen | 11330-32 |
| Heat-inactivated FBS | Hyclone | SH30071.03 |
| B27 supplement | Invitrogen | 17504044 |
| N2 supplement | Invitrogen | 17502048 |
| bFGF | Invitrogen | PHG0026 |
| EGF | R&D system | 236-EG-01M |
| IGF-I | R&D system | 291-G1-050 |
| 0.05% Trypsin/EDTA | Invitrogen | 25300-054 |
| Dispase | Invitrogen | 17105041 |
| Penicillin-Streptomycin | Invitrogen | 15070063 |
Table 1.
References
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