Construction method of diaper dermatitis-like in-vitro recombinant epidermis model

A technology for diaper dermatitis and a construction method, applied in the field of tissue engineering, can solve the problems of lack of diaper-like dermatitis, insufficient accuracy, etc., and achieve the effects of shortening modeling time, reducing individual differences, and avoiding species differences

Active Publication Date: 2020-01-14
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem of the lack of in vitro recombinant epidermal model of diaper-like dermatitis in the prior art and the lack of accuracy of pharmacologic

Method used

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  • Construction method of diaper dermatitis-like in-vitro recombinant epidermis model
  • Construction method of diaper dermatitis-like in-vitro recombinant epidermis model
  • Construction method of diaper dermatitis-like in-vitro recombinant epidermis model

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1: Construction method of the recombinant epidermal model of diaper-like dermatitis

[0038] Step 1. Preparation of Epidermal Cells

[0039] Take primary keratinocytes, recover and expand, use P7 cells, digest with 2.5% trypsin (100 mL PBS, 2.5 g trypsin) digestion solution, use MCDB153 medium to make single cell suspension, follow the 2.5× 10 5 Individual / mL density, inoculated in culture flasks, placed at 37°C, 5% CO 2 cultivated under conditions.

[0040] Step 2, submerged culture of epidermal cells

[0041] Take the cells in the logarithmic growth phase, and use cell culture medium I to make the cell density 2.5×10 6 cells / mL of epidermal cell suspension, inoculate 200 μL into a cell culture chamber containing a polycarbonate filter membrane, and place at 37°C, 5% CO 2 Cultivate under these conditions for 2 hours; then place the small chamber in a model petri dish, add 70 mL of cell culture medium I to each dish, and store at 37°C, 5% CO 2 The culture ...

Embodiment 2

[0067] Example 2: Construction of an in vitro recombinant epidermal model of diaper-like dermatitis

[0068] Step 1. Preparation of Epidermal Cells

[0069] Take the primary keratinocytes, after recovery and expansion, use the P7 generation cells, digest with 2.5% trypsin digestion solution, and make a single cell suspension with MCDB153 medium, according to 5×10 5 Individual / mL density, inoculated in culture flasks, placed at 37°C, 5% CO 2 cultivated under conditions.

[0070] Step 2, submerged culture of epidermal cells

[0071] Cells in the logarithmic growth phase were taken, and the cell density was made 1.25×10 with cell culture medium Ⅰ. 6 200 μL of the epidermal cell suspension per mL was inoculated into a cell culture chamber containing a polycarbonate filter membrane, and placed at 37°C, 5% CO 2 Incubate for 0.5 hours under these conditions; then place the small chamber in a model petri dish, add 80 mL of cell culture medium I to each dish, and store at 37°C, 5% ...

Embodiment 3

[0078] Example 3: Construction of an in vitro recombinant epidermal model of diaper-like dermatitis

[0079] Step 1. Preparation of Epidermal Cells

[0080] Take the primary keratinocytes, after recovery and expansion, use the P7 generation cells, digest with 2.5% trypsin digestion solution, and make a single cell suspension with MCDB153 medium, according to 1×10 5 Individual / mL density, inoculated in culture flasks, placed at 37°C, 5% CO 2 cultivated under conditions.

[0081] Step 2, submerged culture of epidermal cells

[0082] Take the cells in the logarithmic growth phase, and use cell culture medium I to make the cell density 2×10 6 cells / mL epidermal cell suspension, inoculate 200 μL into a cell culture chamber containing a polycarbonate filter membrane, and place at 37°C, 5% CO 2 Cultivate under the same conditions for 2 hours; then place the small chamber in the model culture dish, add 60mL cell culture solution Ⅰ to each dish, 37°C, 5% CO 2 The culture was conti...

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Abstract

The invention particularly relates to a construction method of a diaper dermatitis-like in-vitro recombinant epidermis model. The construction method comprises the steps as follows: step (1): preparation of epidermis cells; step (2) under-solution culture of epidermis cells; step (3) proliferation and differentiation regulation of epidermis cells; and step (4): construction of the diaper dermatitis-like in-vitro recombinant epidermis model. A specific substance is added to a cell culture solution at each stage to regulate proliferation and differentiation of keratinocytes, a barrier weakened in-vitro recombinant epidermis model is obtained, and then, the diaper dermatitis-like in-vitro recombinant epidermis model is constructed by addition of a stimulant. The diaper dermatitis-like in-vitro recombinant epidermis model constructed by the method has structure and biochemical index change similar to tissue slices in a normal pathological state, species difference and individual differenceare reduced, and the real effectiveness of the model is greatly improved.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a method for constructing an in vitro recombined epidermal model of diaper dermatitis. Background technique [0002] Infant skin is an important stage in the transition of skin organs from the amniotic fluid environment of the uterus to the mature state, and its physiological state is significantly different from that of adults. Studies have shown that compared with adult skin, newborn skin has the following characteristics: (1) The stratum corneum and epidermis are thin, the cells are small, the collagen fibers are small and sparse, and the cell density is relatively large, which leads to high skin permeability and It is easy to be damaged by friction, especially the skin folds, which are more likely to cause skin ulcers and diaper rash due to increased friction; (2) The content of natural moisturizing factors is low, the transdermal water loss is more, an...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/0625G01N33/5008G01N33/5044C12N2500/84C12N2501/11C12N2500/40C12N2500/24C12N2501/117C12N2501/39C12N2500/38C12N2501/999C12N2500/14C12N2501/727C12N2503/00Y02A50/30
Inventor 吴迪张勇杰胡成虎何小宁卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
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