US 2014/0227779 A1 · MELANOCYTE OR PROGENITOR CELL THEREOF ADAPTED TO KERATINOCYTE, AND PREPARATION METHOD THEREOF

Abstract

The present invention relates to novel melanocytes and melanoblasts. In addition, the present invention relates to a novel method for producing melanocytes and melanoblasts. Specifically, provided is novel melanocytes of which gene expression, melanin content, and tyrosinase activity are different from those of conventional melanocytes. Even more specifically, provided is novel melanoblasts of which the gene expression, the melanin content the tyrosinase activity, and the protein expression are different from those of conventional melanocytes. Additionally, provided is a novel method for producing melanocytes or melanoblasts by culturing keratinocytes.

Claims22

Dependent claims

A method for producing melanocytes or melanoblasts as progenitor cells thereof adapted to a keratinocyte, which comprises following steps:

(a) culturing keratinocytes in a keratinocyte medium in a dish;
(b) removing the keratinocytes from the culture;
and
(c) collecting cells attached to the bottom of the dish from the culture, in which the keratinocyte is removed, and culturing the collected cells in a melanocyte medium or a melanoblast medium.

1 more element

A melanocyte adapted to a keratinocyte, which has at least one characteristic selected from the following characteristics:

(i) an expression level of p75NTR, which is a neural crest stem cell marker lower than a primary melanocyte;
(ii) an expression level of BRN2, which is expressed in a melanoblast higher than a primary melanocyte;
(iii) a melanin content higher than a primary melanocyte;
(iv) a tyrosinase activity higher than a primary melanocyte;
(v) an increased expression level of p75NTR higher than a primary melanocyte when cultured in a phorbol 12-myristate 13-acetate (PMA)-free medium;
(vi) an increased expression level of BRN2 lower than a primary melanocyte when cultured in a PMA-free medium;
(vii) a characteristic that a relative ratio of a p75NTR expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-free medium, to the p75NTR expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-containing melanocyte medium, is equivalent to 60 to 160% of the ratio of the p75NTR expression level of a primary melanocyte cultured in a PMA-containing melanocyte medium, to the p75NTR expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-containing melanocyte medium;
(viii) a characteristic that a relative ratio of a BRN2 expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-free medium, to the BRN2 expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-containing melanocyte medium, is equivalent to 1 to 10 times of the ratio of the BRN2 expression level of the melanocyte adapted to a keratinocyte cultured in a PMA-containing melanocyte medium, to the BRN2 expression level of the primary melanocyte cultured in a PMA-containing melanocyte medium;
and
(ix) a ratio of the cells attached to the bottom of a dish to the total cultured cells after 2 hour subculture higher than a primary melanocyte.

7 more elements

Dependencies are inferred from the claim text.

Melanocyte or progenitor cell thereof adapted to keratinocyte, and preparation method thereof

Life of this patent

Melanocyte or progenitor cell thereof adapted to keratinocyte, and preparation method thereof

Life of this patent

Abstract

Claims22