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Method for culturing primary cells of solid tumors of bone and soft tissue tumors

A primary cell and cell culture technology, applied in the biological field, can solve the problems of difficult removal of miscellaneous cells, low culture success rate, and long culture cycle, and achieve the effect of high cell purity, high culture stability, and short culture cycle

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. Preparation of reagents for culturing bone and soft tissue tumor solid tumor primary cells

[0076] 1. Sample preservation solution (100mL)

[0077] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0078] Table 1 Sample Preservation Solution (100mL)

[0079]

[0080] 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.

[0081] 2. Sample cleaning solution (100mL)

[0082] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0083] Table 2 Sample cleaning solution (100mL)

[0084]

[0085] The sample cleaning solution should be prepared and used immediately.

[0086] 3. Sample dissociation solution (10mL)

[0087] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0088] Table 3 Sample Dissociation Solution (10mL)

[0089]

...

Embodiment 2

[0159] Example 2. Obtaining of Postoperative Specimens / Biopsy Puncture Specimens of Bone and Soft Tissue Tumor Solid Tumors

[0160] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0161] 2. The attending physician 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 of 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.

[0162] 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 informat...

Embodiment 3

[0164] Example 3, Bone and Soft Tissue Tumor Solid Tumor Tissue Sample Dissociation Pretreatment

[0165] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0166] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0167] 1. Sample weighing.

[0168] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0169] 3. Wash the sample 5 times with sample cleaning solution, and wash the sample 5 times with sterile PBS solution.

[0170] 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 method for culturing primary cells of solid tumors of bone and soft tissue tumors. The invention provides a method for culturing primary cells of solid tumors of bone and soft tissue tumors and a matched reagent. According to the technical core, a mild cell dissociation reagent is used for treating bone and soft tissue tumor solid tumor tissues, so that the activity of tumor cells in the tissues is ensured to the greatest extent; and a special serum-free culture medium is prepared, solid tumor cells from bone and soft tissue tumors are subjected to in-vitro culture by using a suspension culture system, and interference of normal cells is eliminated to the greatest extent while normal amplification of the tumor cells is 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 method for culturing primary cells of solid tumors of bone and soft tissue tumors. 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. [0003] ...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01N1/02
CPCC12N5/0693A01N1/0226C12N2500/32C12N2501/115C12N2501/12C12N2501/105C12N2501/13C12N2500/25C12N2501/998C12N2501/727C12N2500/46C12N2501/392C12N2509/00
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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