Differentiation of adipose-derived stem cells to functional CD105(neg) CD73(low) melanocyte precursors guided by defined culture condition

Zavala G.; Sandoval C.; Meza D.; Contreras R.; Gubelin W.; Khoury M.

Abstract

Background The generation of functional human epidermal melanocytes (HEM) from stem cells provides an unprecedented source for cell-based therapy in vitiligo. Despite the important efforts exerted to obtain melanin-producing cells from stem cells, pre-clinical results still lack the safety and scalability characteristics essential for their translational application. Methods Here, we report a rapid and efficient protocol based on defined culture conditions capable of differentiating adult adipose-derived stem cells (ADSC) to scalable amounts of proliferative melanocyte precursors (PreMel) within 30 days. PreMel were characterized in vitro through qPCR, Western blot, flow cytometry, biochemical assays, and in vivo assays in immunocompromised mice (NOD.Cg-Prkdc(scid) Il2rg(tm1Wjl)/SzJ, or NSG). Results After 30 days of differentiation, the stem cell-derived PreMel were defined as CD105(neg) CD73(low) according to immunophenotypic changes in comparison with parental stem cell markers. In addition, expression of microphthalmia-associated transcription factor (MITF), active tyrosinase (TYR), and the terminal differentiation-involved premelanosome protein (PMEL) were detected. Furthermore, PreMel had the potential to synthesize melanin and package it into melanosomes both in vitro and in vivo in NSG mice skin. Conclusions This study proposes a rapid and scalable protocol for the generation of proliferative melanocyte precursors (PreMel) from ADSC. These PreMel display the essential functional characteristics of bona fide HEM, opening a new path for an autologous cellular therapy for vitiligo patients.

Más información

Título según WOS: Differentiation of adipose-derived stem cells to functional CD105(neg) CD73(low) melanocyte precursors guided by defined culture condition
Título según SCOPUS: Differentiation of adipose-derived stem cells to functional CD105neg CD73low melanocyte precursors guided by defined culture condition
Título de la Revista: STEM CELL RESEARCH & THERAPY
Volumen: 10
Número: 1
Editorial: BMC
Fecha de publicación: 2019
Idioma: English
DOI:

10.1186/s13287-019-1364-0

Notas: ISI, SCOPUS