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Culture medium special for organoids of nasopharyngeal carcinoma and scaffold-free culture method

A nasopharyngeal carcinoma, culture medium technology, applied in tissue culture, 3D culture, cell culture active agent, etc., can solve the problems of poor cell growth, difficult removal of scaffold materials, etc., to improve the culture speed and success rate, uniform size, The effect of shape rules

Active Publication Date: 2020-09-22
江苏信安佳医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a special culture medium for nasopharyngeal carcinoma that does not require scaffold materials, and also provides a supporting The unique culture method can quickly form organoids with uniform size, regular shape, and stable genetic and biological properties

Method used

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  • Culture medium special for organoids of nasopharyngeal carcinoma and scaffold-free culture method
  • Culture medium special for organoids of nasopharyngeal carcinoma and scaffold-free culture method
  • Culture medium special for organoids of nasopharyngeal carcinoma and scaffold-free culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Nasopharyngeal carcinoma organoid-specific medium formula:

[0052] EGF 20ng / mL, Noggin 200ng / mL, Y-27632 10μM, A83-01 0.6μM, SB202190 10μM, bFGF 5ng / mL, hydrocortisone 80ng / mL, Insulin 1μg / mL, penicillin / streptomycin double antibody 1 %, FBS 2%, Keratinocyte-SFM added to the required volume.

Embodiment 2

[0054] Tissue washing solution formula: PBS solution containing penicillin / streptomycin 500U / mL and amphotericin B 12.5mg / L.

Embodiment 3

[0056] Tissue digestion solution formula: Hank's solution containing 0.1% (mass percentage) of type I collagen and 0.1% (mass percentage content) of type IV collagen.

[0057] Example 4 Nasopharyngeal Carcinoma Tissue Separation

[0058] 1. Fresh cancer tissue processing

[0059] The fresh tumor tissue of nasopharyngeal carcinoma was cleaned with 4°C pre-cooled tissue washing solution. Cut the tissue into small pieces with sterilized ophthalmic scissors in a centrifuge tube, the size of the tissue piece is 1mm 2 about. After adding 5 mL of tissue digestion solution (about 10 times the volume), place the centrifuge tube in a shaker in a 37°C incubator and digest for 30 minutes.

[0060] The cells in the digestion solution were filtered with a 200-mesh nylon mesh, and 5 mL of Keratinocyte-SFM medium containing 5% FBS was added to the filtrate to terminate the digestion. The remaining tissue was added to the tissue digestion solution for secondary digestion, and the filtrate ...

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Abstract

The invention discloses a culture medium special for organoids of nasopharyngeal carcinoma and a scaffold-free culture method. The culture medium comprises EGF, Noggin, Y-27632, A83-01, SB202190, bFGF, hydrocortisone, Insulin, penicillin-streptomycin, FBS and Keratinocyte-SFM. The culture method comprises the following steps: inoculating separated tumor cells of nasopharyngeal carcinoma into an ultra-low adsorption U-shaped cell culture plate, adding the culture medium special for organoids of nasopharyngeal carcinoma, performing centrifugation, performing culture in a CO2 incubator with a volume concentration of 5% at a temperature of 37 DEG C, and replacing a scaffold-free culture medium once every 3-4 days. According to the culture medium special for organoids of nasopharyngeal carcinoma and the scaffold-free culture method, introduction of an exogenous scaffold material is not needed, cell aggregation and clustering can be promoted under the assistance of gravity, rapid amplification of the tumor cells in a short time is achieved so as to construct the organoids, the success rate can reach 100%, and the specificity of primary tumor can be maintained; and meanwhile, the activityand functional and high expression of the tumor cells can be maintained for a long period of time.

Description

technical field [0001] The invention relates to the field of tumor organoid culture, and further relates to the field of nasopharyngeal carcinoma organoid culture, in particular to a special culture medium for nasopharyngeal carcinoma organoid and a scaffold-free culture method. Background technique [0002] Nasopharyngeal carcinoma is derived from nasopharyngeal epithelial cells and is a malignant tumor of the epithelial tissue of the nasopharynx. It has obvious regional characteristics in epidemiology, and its incidence is high in southern my country and Southeast Asia. Nasopharyngeal carcinoma has different pathogenic factors and tumor biological behaviors from other head and neck squamous cell carcinomas. With the widespread application of induction chemotherapy, concurrent chemoradiotherapy and intensity-modulated radiotherapy in the treatment of nasopharyngeal carcinoma, the local control rate of nasopharyngeal carcinoma has been significantly improved. However, local...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071
CPCC12N5/0693C12N5/0697C12N2513/00C12N2501/11C12N2501/115C12N2501/30C12N2501/33C12N2501/999
Inventor 罗国安王义明范雪梅罗喆明
Owner 江苏信安佳医疗科技有限公司
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