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Method for cryopreserving tumor tissue

A tumor tissue, cryopreservation technology, applied in the preservation, application, animal husbandry and other directions of human or animal body, to achieve the effect of high tumor formation rate

Inactive Publication Date: 2016-08-17
SHAOXING PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, there are few reports on how to cryopreserve the tumor tissue of a tumor patient so that the transplanted tumor can relatively completely retain the morphological and molecular characteristics of the patient-derived tissue, and at the same time have a higher tumor formation rate

Method used

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  • Method for cryopreserving tumor tissue
  • Method for cryopreserving tumor tissue
  • Method for cryopreserving tumor tissue

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1: Obtaining and Freezing of Patient Tumor Tissue Specimens

[0022] Fresh tumor tissue samples were obtained from a 40-year-old female patient with colon cancer and lymph node and liver metastases. The samples were obtained with the informed consent of the patient and the permission of the ethics committee of the institution. The patients had not received chemotherapy and other treatments before operation. The intraoperative specimens were cut into different sizes with sterilized surgical scissors and then frozen in the pre-prepared freezing solution of the present invention (first placed in a freezing box containing isopropanol at 4°C for about 1 hour; Then place the frozen storage box containing the specimen in a low-temperature refrigerator at -20°C overnight (24 hours); then place the frozen storage box containing the specimen at -80°C (24 hours); and remove the specimen from the freezing storage box Take it out and place it in liquid nitrogen (-196°C) for...

Embodiment 2

[0026] Example 2: Establishment of multi-focal PDTT xenograft model of colon cancer

[0027] 1) BALB / c nude mice aged 4-6 weeks were raised in an SPF barrier environment and allowed to adapt to the environment for more than three days;

[0028] 2) The tumor tissue specimens frozen in the cryopreservation solution of the present invention (the specimens have been cut into fragments of different sizes with sterilized surgical scissors before freezing) are recovered and transferred to the RPMI 1640 medium (containing 20% fetal bovine serum and 0.05% penicillin / streptomycin);

[0029] 3) Then wash three times with the above-mentioned RPMI 1640 medium;

[0030]4) After the animal is anesthetized with isoflurane, disinfect the skin on the back and abdomen of the nude mouse, wipe it dry with a sterile cotton ball, cut a small incision near the anterior armpit on one side of the nude mouse with a razor blade, and bluntly separate the skin and subcutaneous tissue with a 18-gauge punct...

Embodiment 3

[0035] Example 3: Effects of Different Freezing Time on the Tumor Formation Rate of Colon Cancer PDTT Transplanted Tumor Model

[0036] According to the method of Example 2, the PDTT xenograft tumor model was established according to the tumor tissue with the cut size of 1 mm×1 mm×3 mm frozen for 2-52 weeks according to Example 1, and the tumor formation rate is shown in Table 1.

[0037] Table 1: The effect of different cryopreservation time on the tumor formation rate of colon cancer PDTT xenograft model

[0038]

[0039]

[0040] Note: G3, passage to the third generation.

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Abstract

The invention relates to a method for cryopreserving tumor tissue. The method includes the steps that tumor tissue from a tumor patient is sheared to be in the size of 1 mm+ / -0.2*1+ / -0.2 mm*3+ / -0.6 mm, and then the sheared tissue blocks are cryopreserved for 25+ / -2 weeks. Optionally, the tissue cryopreserved for 25+ / -2 weeks are used for building a PDTT transplantation tumor model or in-vivo or in-vitro serial subcultivation. The cryopreserving method has the advantages that the tumor formation rate is high, and the form and the molecular characteristic of source tissue of the patient can be completely reserved through transplantation tumor.

Description

Technical field: [0001] The invention belongs to the field related to the preservation of biological tissues or cells, and relates to a method for freezing tumor tissues. Background technique: [0002] In the process of drug development and development of specific therapeutic approaches or medications, biological tissues (such as biological tissues obtained by surgical resection or puncture, especially tumor tissues) need to be preserved for subsequent treatment, detection or scientific research etc. At present, the commonly used method of preserving biological tissues is cryopreservation, and cryopreservation also has a series of effects on biological tissues. For example, Zhang JN, Yu YY, Ji J, et al.Effect of time offreeze-preserving on biological macromolecules of tumor tissue[J].Journal ofDiagnostics Concepts&Practice, 2009,8(1):38-42 discusses the effect of cryopreservation time on biological macromolecules of tumor tissue. The influence of biological macromolecules ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0263
Inventor 陶锋金科涛蓝欢荣黄黎明吕杰青景元明王伟胡耕远
Owner SHAOXING PEOPLES HOSPITAL
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