Method for directly reprogramming mouse embryo fibroblasts into melanocytes

Abstract

The invention provides a method for directly reprogramming mouse embryo fibroblasts into melanocytes. The method comprises the following steps: preparing high-quality concentrated lentivirus for transfecting alternative transcription factors; directly reprogramming the mouse embryo fibroblasts into melanocytes; optimizing and screening transcription factors capable of being used for direct reprogramming of melanocytes; and identifying the function of inducing melanocytes. The invention relates to an optimized and efficient melanocyte direct reprogramming method and a simplified and optimized melanocyte direct reprogramming system, and the method mainly comprise two aspects: preparing high-quality concentrated viruses, optimizing reprogramming culture medium components, and improving functions of melanocytes obtained by direct reprogramming; screening the transcription factor with the most significant effect for direct reprogramming of melanocytes; carrying out functional identification on the directly reprogrammed melanocytes, and providing a new treatment strategy for patients suffering from depigmentation diseases such as vitiligo.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a method for directly reprogramming mouse embryonic fibroblasts into melanocytes. Background technique [0002] Vitiligo is a kind of skin disease that seriously affects people's physical and mental health. Due to gene mutation, oxidative stress, autoimmunity, metabolic disorders and other reasons, the lack of melanocytes stops the secretion of melanin, and then the vitiligo disease occurs. In particular, the large area of ​​vitiligo that grows on the face has brought great distress to patients (Ezzedine K, EleftheriadouV, Whitton M, van Geel N. Vitiligo. Lancet. 2015 Jul 4; 386(9988): 74-84.doi : 10.1016 / S0140-6736 (14) 60763-7. Epub 2015 Jan 15. PMID: 25596811.). Currently, corticosteroids, immunomodulators and other drugs can be used for the treatment of vitiligo (Speeckaert R, van Geel N. Vitiligo: An Update on Pathophysiology and Treatment Options. Am J Cl...

AI Technical Summary

Problems solved by technology

However, because these combinations contain a variety of transcription factors, and the final results are not consistent, a small number of transcription factors have not been identified, so for a specific transcription factor, its role is not yet clear, and it is also difficult to make a decision in the process of direct melanin reprogramming

[0008] Complicated transcription factors also lead to the complexity of these technical methods, decreased cell viability, low transdifferentiation efficiency, poor functionality of the obtained cells, and no way to compare with positive references

In addition, the composition of melanin direct reprogramming medium is not clear, and researchers need a lot of time to explore, which not only increases the cost of the experiment, but also prolongs the time of the experiment, and is not conducive to the subsequent large-scale application

[0009] Although cell transdifferentiation is a hot research direction at present, there are not many studies on the direct reprogramming of melanocytes and there is no conclusion on the method of melanocyte reprogramming, whether it is transcription factors or differentiation promotion in culture medium. The choice of factors is not clear, the system of direct reprogramming is complex and inefficient, and generally the cells derived from direct reprogramming are poorly functional, far from the positive reference comparison

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