Mixed cultivation method of keratinocyte and melanocyte and application
A technology of keratinocytes and melanocytes, applied in the field of mixed culture of keratinocytes and melanocytes, which can solve the problems of inefficiency, shortage of skin sources for patients, poor uniformity, etc., and achieve the effect of avoiding pollution
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example 1
[0052] Example 1: Successful isolation and expansion of human epidermal cells from adult normal breast skin
[0053] (1) Materials and methods
[0054] Sample: adult normal chest skin (including epidermis and dermis) discarded during hospital surgery, about 3cm 2 .
[0055] Skin Media: KGM-Gold vs. KGM
[0056] (2) Cell isolation and skin slice culture
[0057] a) The skin samples were transported to the laboratory in refrigerated DMEM medium, and the area was measured.
[0058] b) Wash the sample with PBS; sterilize with iodine and alcohol and then wash the sample again, remove the yellow fat layer and most of the white dermis in the tissue slice, keep the epidermis and 0.2-3mm thick dermis, and decompose the skin slice into tiny particles .
[0059] c) 0.05% trypsin cold digestion sample for 12-18 hours; add trypsin stop solution; filter to collect epidermal tissue.
[0060] d) Add type I collagenase and magnetically stir for 2-6 hours to heat digest, filter, centr...
example 2
[0068] Example 2: The foreskin skin of a healthy male after circumcision was successfully isolated and expanded to culture human epidermal cells
[0069] (1) Materials and methods
[0070] Sample: Foreskin skin (including epidermis and dermis) of healthy males after circumcision, about 4cm 2 .
[0071] Skin Media: KGM-Gold vs. KGM
[0072] (2) Cell isolation and skin slice culture
[0073] a) The skin samples were transported to the laboratory in refrigerated DMEM medium, and the area was measured.
[0074] b) Wash the sample with PBS; sterilize with iodine and alcohol and then wash the sample again, remove the yellow fat layer and most of the white dermis in the tissue slice, keep the epidermis and 1-3mm thick dermis, and decompose the skin slice into tiny particles .
[0075] c) 0.05% trypsin cold digestion sample for 15 hours; add trypsin stop solution; filter to collect epidermal tissue.
[0076] d) adding type I collagenase and magnetic stirring for 5 hours of t...
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