Method for analyzing toxicity and effect of skin whitening agent through human being skin melanocyte
A technology of melanocytes and whitening agent, applied in the field of toxicology and efficacy experiments, can solve the problems of high cost, immaturity, and unstable melanin expression.
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Embodiment 1
[0061] I. Culture and Identification of Human Skin Primary Melanocytes
[0062] (1) Configuration of human skin melanocyte culture medium
[0063] 1). Preparation of human melanocyte culture medium:
[0064]A. Complete culture solution: Mix keratinocyte serum-free medium K-SFM (Keratinocyte Serum-Free Medium) with fetal calf serum at a volume ratio of 9:1, and add the following cofactors: Hydrocortisone (Hydrocortisone), and its final The concentration is 0.4ug / ML; bovine insulin (insuilin), its final concentration is 10ug / ML; L-glutamine (L-glutamine), its final concentration is 6mmol; 12-o-tetradecanoyl phorbol acetate -13 (abbreviated as TPA, English name 12-o-tetradecanoylphorbol-13-acetate), its final concentration is 81.06nmol / L; 3-isobutyl-1-methylxanthine (abbreviated as IBMX, namely 3-isobutyl- l-methylxanthine), its final concentration is 0.1nmol / L; transferrin (transferrin), its final concentration is 10ug / ML; cholera toxin (cholera toxin, CT), its final concentra...
Embodiment 2
[0093] I. Culture and Identification of Human Skin Primary Melanocytes
[0094] (1) Configuration of human skin melanocyte culture medium
[0095] 1) Preparation of human melanocyte culture medium:
[0096] A. Complete culture solution: Mix keratinocyte serum-free medium K-SFM (Keratinocyte Serum-Free Medium) with fetal bovine serum at a volume ratio of 9:1, and add the following cofactors: hydrocortisone (Hydrocortisone), and its final Concentration is 0.4ug / ML; Bovine insulin (insuilin), its final concentration is 10ug / ML; L-glutamine (L-glutamine), its final concentration is 6mmol; 12-o-tetradecanoyl phorbol acetate -13 (abbreviated as TPA, English name 12-o-tetradecanoylphorbol-13-acetate), its final concentration is 81.06nmol / L; 3-isobutyl-1-methylxanthine (abbreviated as IBMX, namely 3-isobutyl- l-methylxanthine), its final concentration is 0.1nmol / L; transferrin (transferrin), its final concentration is 10ug / ML; cholera toxin (choleratoxin, CT), its final concentratio...
Embodiment 3
[0125] I. Culture and Identification of Human Skin Primary Melanocytes
[0126] (1) Configuration of human skin melanocyte culture medium
[0127] 1) Preparation of human melanocyte culture medium:
[0128] A. Complete culture solution: Mix keratinocyte serum-free medium K-SFM (Keratinocyte Serum-Free Medium) with fetal bovine serum at a volume ratio of 9:1, and add the following cofactors: hydrocortisone (Hydrocortisone), and its final The concentration is 0.4ug / ML; bovine insulin (insuilin), its final concentration is 10ug / ML; L-glutamine (L-glutamine), its final concentration is 6mmol; 12-o-tetradecanoyl phorbol acetate -13 (abbreviated as TPA, English name 12-o-tetradecanoylphorbol-13-acetate), its final concentration is 81.06nmol / L; 3-isobutyl-1-methylxanthine (abbreviated as IBMX, namely 3-isobutyl- l-methylxanthine), its final concentration is 0.1nmol / L; transferrin (transferrin), its final concentration is 10ug / ML; cholera toxin (choleratoxin, CT), its final concentr...
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