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Methods for preparing human melanocytes from human pluripotent stem cells

A technology of pluripotent stem cells and melanocytes, applied in non-embryonic pluripotent stem cells, epidermal cells/skin cells, biochemical equipment and methods, etc.

Inactive Publication Date: 2013-01-02
INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, a method for generating human melanocytes from human pluripotent stem cells using a 2D protocol has not been reported in the prior art

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  • Methods for preparing human melanocytes from human pluripotent stem cells

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specific Embodiment approach

[0146] Method for producing melanocyte populations and human skin substitutes from human pluripotent stem cells

[0147] In this report, the inventors demonstrate that iPS reprogrammed cells (hiPSCs) and hESCs are capable of generating pure and functional melanocyte populations. These cells were obtained by following a directed 2D protocol for epidermal induction of trophoblast cells in FAD medium supplemented with BMP4 and ascorbic acid. These melanocytes, derived from pluripotent stem cells, express all major markers of melanocytes, develop melanosomes and synthesize melanin that can migrate into keratinocytes.

[0148] Materials and Methods

[0149] hESC culture and melanocyte differentiation

[0150] From two cell lines SA01 (Cellartis, Sweden) and WA09 (Wicell, Madison WI) hESCs and hiPSCs produced using retroviral Yamanaka factors (Takahashi et al.) and lentiviral Thomson factor (Thomsom factor) (Thomson et al.) Mitomycin C inactivated STO mouse fibroblasts were gro...

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Abstract

The invention relates to an ex vivo method for obtaining a population of human melanocytes derived from human pluripotent stem cells comprising the step consisting of co-culturing human pluripotent stem cells with cells that support ectodermal differentiation in the presence of an agent that stimulates epidermal induction and an agent that stimulates terminal differentiation of keratinocytes. The invention also relates to human melanocytes obtainable by said method and to uses thereof in cell therapy and in screening assays.

Description

technical field [0001] The present invention relates to an ex vivo method for obtaining a population of human melanocytes derived from human pluripotent stem cells. Background technique [0002] Melanocytes are pigment producing cells responsible for the coloring of the skin, eyes and hair. In vertebrate development, melanocytes originate from the neural crest and ultimately reside in the epidermis after undergoing a complex process of fate specification, proliferation, migration, survival and differentiation. Pigmentation results from the highly controlled production of melanin in specialized organelles called melanosomes. Through a process called melanogenesis, melanosomes are transported along the dendrites and transported to growing hair, or to surrounding keratinocytes, where they play a role in protecting human tissue against DNA damage and tissue cancer play an important role in the harmful effects of sunlight ultraviolet (UV) radiation. Over the past 10 years, sev...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
CPCC12N2500/40C12N2501/01C12N2501/155C12N2501/33C12N2501/11C12N2506/02C12N2500/38C12N5/0626C12N2501/39C12N2502/16C12N2501/395C12N2502/13A61P17/00C12N5/00C12N5/0606C12N5/0607
Inventor X·尼桑C·巴尔德希G·勒迈特M·佩尚斯基
Owner INST NAT DE LA SANTE & DE LA RECHERCHE MEDICALE (INSERM)
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