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Culture medium for culturing primary cells of solid tumors of bone and soft tissue tumors

A technology of primary cells and culture medium, applied in tumor/cancer cell, culture process, tissue culture, etc., can solve the problems of low success rate of culture, long culture period, difficult to remove foreign cells, etc., to achieve high cell purity and culture. The effect of short cycle and less interference of heterocysts

Active Publication Date: 2021-05-07
SUZHOU GENOARRAY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing primary tumor cell culture technologies mainly include 2D culture, 3D culture, reprogramming culture, etc. These methods face the problems of extremely long culture cycle, low culture success rate, and difficulty in removing stray cells to varying degrees.

Method used

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  • Culture medium for culturing primary cells of solid tumors of bone and soft tissue tumors
  • Culture medium for culturing primary cells of solid tumors of bone and soft tissue tumors
  • Culture medium for culturing primary cells of solid tumors of bone and soft tissue tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1. Preparation of reagents for culturing bone and soft tissue tumor solid tumor primary cells

[0052] 1. Sample preservation solution (100mL)

[0053] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0054] Table 1 Sample Preservation Solution (100mL)

[0055]

[0056] After the sample preservation solution is prepared, it is divided into 15mL centrifuge tubes, 5mL per tube. After aliquoting, it can be stored at 4°C for 1 month.

[0057] 2. Sample cleaning solution (100mL)

[0058] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0059] Table 2 Sample cleaning solution (100mL)

[0060]

[0061] The sample cleaning solution should be prepared and used immediately.

[0062] 3. Sample dissociation solution (10mL)

[0063] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0064]Table 3 Sample Dissociation Solution (10mL)

[0065] ...

Embodiment 2

[0135] Example 2. Obtaining of Postoperative Specimens / Biopsy Puncture Specimens of Bone and Soft Tissue Tumor Solid Tumors

[0136] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0137] 2. The attending doctor selects the patients according to the clinical indications stipulated in the medical guidelines, and selects appropriate samples for in vitro culture according to the clinical indications during the operation. The selection criteria for the samples are: primary osteosarcoma, synovial sarcoma, Rhabdomyosarcoma, Leiomyosarcoma, Ewing's Sarcoma, Liposarcoma, Acinar Soft Tissue Sarcoma, Clear Cell Sarcoma, Fibroma, Surgical specimens weighing more than 20 mg.

[0138] 3. The attending doctor provides basic clinical information such as the patient's gender, age, medical history, family history, smoking history, pathological staging and typing, and clinical diagnosis. The patient's name, ID number and other informatio...

Embodiment 3

[0140] Example 3, Bone and Soft Tissue Tumor Solid Tumor Tissue Sample Dissociation Pretreatment

[0141] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0142] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0143] 1. Sample weighing.

[0144] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0145]3. Wash the sample 5 times with sample cleaning solution, and wash the sample 5 times with sterile PBS solution.

[0146] 4. Use ophthalmic scissors, ophthalmic forceps, scalpel and other equipment to carefully peel off the adipose tissue, connective tissue, and necrotic tissue in the sample.

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Abstract

The invention discloses a culture medium for culturing primary cells of solid tumors of bone and soft tissue tumors. The culture medium for culturing the primary cells of the solid tumors of the bone and soft tissue tumors provided by the invention is a special serum-free culture medium, and the culture medium is used for carrying out in-vitro suspension culture on solid tumor cells derived from the bone and soft tissue tumors, so that the interference of normal cells can be eliminated to the greatest extent while normal amplification of the tumor cells can be ensured. The bone and soft tissue tumor solid tumor primary cell culture obtained by the method can be used for various cellular in-vitro experiments, next-generation sequencing, animal model construction, cell line construction and the like. Predictably, the culture method has a wide application prospect in the fields of research and clinical diagnosis and treatment of solid tumors of bone and soft tissue tumors.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a culture method base for culturing bone and soft tissue tumor solid tumor primary cells. Background technique [0002] Bone and soft tissue tumors are diseases that seriously endanger human health and life. In recent years, the incidence rate has gradually increased. Primary malignant bone tumors are more common in adolescents and middle-aged people, and the common ones are osteosarcoma, Ewing sarcoma, chondrosarcoma, and malignant fibrous histiocytic cells. tumors, chordomas, etc. Common soft tissue malignancies are synovial sarcoma, fibrosarcoma, liposarcoma, rhabdomyosarcoma, etc. The incidence of bone and soft tissue tumors accounts for about 1% of the incidence of malignant tumors, but there are many types of them, resulting in non-standard diagnosis and treatment. Current clinical guidelines recommend the use of clinical trials for the treatment of soft tissue sarcomas. [0...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/105C12N2500/46C12N2501/392C12N2500/90C12N2501/115C12N2501/12C12N2500/25C12N2501/13C12N2500/60C12N2500/32C12N2501/727Y02A50/30
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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