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Oral mucosa epithelium model and in-vitro construction method thereof

A technology of oral mucosa epithelium and oral mucosa, which is applied in biochemical equipment and methods, microbial determination/inspection, tumor/cancer cells, etc. Application and other problems, to achieve the effect of solving the difficulty of seed cell source, stable and controllable cell line, and improving barrier function

Inactive Publication Date: 2016-01-20
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, HanneM?rckNielsen et al. used TR146 cells to construct an oral mucosa model in vitro for research, but this model has a series of defects: First, the construction time is long, and it takes up to one month to construct a batch of models in vitro, and excessive construction time will increase Uncontrollable factors in the process affect the stability of the model; second, the single culture system cannot adapt to the nutritional needs of cells at different stages, resulting in poor stratification of the model, no hemidesmosome structure in the basal layer, uneven and complete structure, and poor barrier function. Weakened, seriously reducing the application of the model; 3, mainly used in the study of drug penetration and absorption, with a single application direction

Method used

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  • Oral mucosa epithelium model and in-vitro construction method thereof
  • Oral mucosa epithelium model and in-vitro construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example focuses on the in vitro construction method steps of oral mucosal squamous carcinoma cell line TR146 as oral mucosal cells:

[0032] Step 1. Expansion and culture of oral mucosal squamous carcinoma cell TR146:

[0033] Oral mucosal squamous carcinoma cell line TR146 was taken, and after recovery and expansion, the P10 generation cells were used, digested with trypsin to make a single-cell suspension, and the cell density was adjusted to 5.0×10 5 Cells / mL, inoculated in a culture bottle, cultured in culture solution I with 10% fetal bovine serum, gently shake the culture bottle to make the cells disperse evenly, and place at 37°C, 5% CO 2 cultivated under conditions;

[0034] The culture solution I is DMEM, which contains 4.5 g / L of glucose and 2.0 mM of glutamine;

[0035] Step 2. Inoculation and culture of oral mucosal epithelial cells under liquid

[0036] Oral mucosal cells in the logarithmic growth phase were taken, and single-cell suspension was mad...

Embodiment 2

[0048] This example focuses on the in vitro construction method steps of oral mucosal squamous carcinoma cell line SCC-25 as oral mucosal cells:

[0049] Step 1. Expansion and culture of oral mucosal squamous carcinoma cell line SCC-25:

[0050] The oral mucosal squamous carcinoma cell line SCC-25 was taken, and after recovery and expansion, the P10 generation cells were used, digested with trypsin to make a single-cell suspension, and the cell density was adjusted to 5.0×10 5 Cells / mL, inoculated in a culture bottle, cultured in culture solution I with 10% fetal bovine serum, gently shake the culture bottle to make the cells disperse evenly, and place at 37°C, 5% CO 2 incubator culture;

[0051] The culture solution I is DMEM, which contains 4.5 g / L of glucose and 2.0 mM of glutamine;

[0052] Step 2. Inoculation and culture of oral mucosal epithelial cells under liquid

[0053] Take the cells in the logarithmic growth phase, make a single cell suspension with culture medium...

Embodiment 3

[0061] This example focuses on the in vitro construction method steps of oral mucosal squamous carcinoma cell lines HN-6 and CAL-27 as oral mucosal cells:

[0062] Step 1. Expansion and culture of oral mucosal squamous carcinoma cells HN-6 and CAL-27:

[0063] Oral mucosal squamous carcinoma cell lines HN-6 and CAL-27 were recovered and amplified respectively. Cells of the P10 generation were digested with trypsin to make a single-cell suspension, and the cell density was adjusted to 5.0×10 5 cells / mL, inoculated in culture flasks respectively, cultured in culture solution Ⅰ with 10% fetal bovine serum, gently shake the culture flasks to disperse the cells evenly, and place at 37°C, 5% CO 2 incubator culture;

[0064] The culture solution I is DMEM, which contains 4.5 g / L of glucose and 2.0 mM of glutamine;

[0065] Step 2. Inoculation and culture of oral mucosal epithelial cells under liquid

[0066] Take HN-6 and CAL-27 cells in the logarithmic growth phase respectively, ...

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Abstract

The invention provides an oral mucosa epithelium model and an in-vitro construction method thereof and belongs to the technical field of tissue engineering biological materials. The in-vitro oral mucosa epithelium model is formed through stratification of one or two of oral mucosa squamous cancer cell lines TR146, Tca8113, SCC-4, SCC-9, SCC-25,HN-6 and CAL-27 with an air-liquid interface culture method adopting different culture systems at different stages after in-vitro amplification according to cytobiology and engineering principles.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering biomaterials, and specifically relates to the safety and efficacy of oral care products, small molecular compounds, active proteins, medical devices, chemicals, disposable hygiene products and other products that can replace oral mucosal epithelium in vitro Oral mucosal epithelial model and its construction method for safety testing and other materials in contact with oral mucosal epithelium for safety and efficacy evaluation. Background technique [0002] The epithelial model of the oral mucosa in the present invention is constructed by applying the principles of cell biology and engineering, and a small amount of seed cells are amplified in vitro. layer, spinous layer and basal layer, all of which have no cuticle and have a complete epithelial structure. In addition to being used to replace or repair diseased and defective tissues and reconstruct physiological functions, the reconstru...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
Inventor 何欣卢永波李潇
Owner GUANGDONG BOXI BIO TECH CO LTD
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