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A new tissue cryopreservation solution

A cryopreservation solution and cryopreservation technology, applied in the preservation, application, animal husbandry and other directions of human or animal bodies, can solve the problems of short cryopreservation time and low tumor formation rate, and achieve a simple preparation method and high tumor formation rate. , can be frozen for a long time

Inactive Publication Date: 2018-08-17
SHAOXING PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Existing tumor tissue cryopreservation solutions (for example, containing 10% DMSO, 40% calf serum, 50% RPMI 1640) have defects such as low tumor formation rate and short freezing storage time, so there is an urgent need for a high tumor formation rate, cryopreservation solution, etc. Tumor tissue cryopreservation solution for a long time

Method used

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  • A new tissue cryopreservation solution
  • A new tissue cryopreservation solution
  • A new tissue cryopreservation solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Obtaining and Freezing of Patient Tumor Tissue Specimens

[0019] 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. Cut the intraoperative specimen into 1×1×3mm with sterilized surgical scissors 3 fragments of different sizes and frozen in the pre-prepared cryopreservation solution of the present invention (first placed in a freezer box containing isopropanol at 4°C for about 1 hour; then put the freezer box containing the specimen into Overnight in a low-temperature refrigerator at -20°C (24 hours); then place the freezer box containing the specimen at -80°C (24 hours); and take the specimen out of the freezer box and place it in liquid nitrogen (-196°C) long-term stora...

Embodiment 2

[0020] Example 2: Establishment of Multifocal PDTT Transplanted Tumor Model of Colon Cancer

[0021] 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;

[0022] 2) transfer the tumor tissue specimen frozen in the freezing solution of the present invention according to Example 1 to ice-bathed RPMI 1640 medium (containing 20% ​​fetal bovine serum and 0.05% penicillin / streptomycin);

[0023] 3) wash three times with the above-mentioned RPMI 1640 medium;

[0024] 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 puncture needle. , make it into a leather bag, place the prepared tumor fragments in it, add a drop of double ant...

Embodiment 3

[0029] Example 3: Freezing solution formula and its related tumor formation rate

[0030] Recipe 1:

[0031] Leibovitz L-15 Medium

[0032] 5% (volume percentage, the same below) fetal bovine serum

[0033] 5% (volume percentage, the same below) dimethyl sulfoxide (DMSO)

[0034] Recipe 2:

[0035] Leibovitz L-15 Medium

[0036] 10% fetal bovine serum

[0037] 10% Dimethyl Sulfoxide (DMSO)

[0038] Recipe 3:

[0039] Leibovitz L-15 Medium

[0040] 15% fetal bovine serum

[0041] 15% DMSO (dimethyl sulfoxide)

[0042] Recipe 4:

[0043] Leibovitz L-15 Medium

[0044] 20% fetal bovine serum

[0045] 20% DMSO (dimethyl sulfoxide)

[0046] Recipe 5:

[0047] Leibovitz L-15 Medium

[0048] 30% fetal bovine serum

[0049] 20% DMSO (dimethyl sulfoxide)

[0050] Recipe 6:

[0051] Leibovitz L-15 Medium

[0052] 14% Fetal Bovine Serum

[0053] 14% DMSO (dimethyl sulfoxide)

[0054] Recipe 7:

[0055] Leibovitz L-15 Medium

[0056] 14% Fetal Bovine Serum

[0057]...

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Abstract

The invention belongs to the field related to biological tissue or cell preservation, and relates to a novel tissue cryopreservation liquid, in particular to a liquid used for cryopreserving tumor tissue or ovarian tissue. The cryopreservation liquid is prepared from a Leibovitz L-15 culture medium, fetal calf serum and 14-18% of DMSO (dimethyl sulfoxide), wherein the Leibovitz L-15 culture medium (Gibco) can be purchased or can be self-made by technicians in the field according to a reported formula; preferably, the content of DMSO is 14-16%, more preferably, 15%. For the novel cryopreservation liquid, preferably, the content of fetal calf serum is 14-18%, more preferably, 14-16%, the most preferably, 15%. The cryopreservation liquid has the advantages of being long in cryopreservation time, high in tumor formation rate and the like, the preparation method is simple, and the raw material source is wide.

Description

Technical field: [0001] The invention belongs to the field related to the preservation of biological tissues or cells, and relates to a liquid for freezing tumor tissues or ovarian 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. For example: Chinese patent application CN201510261052.9 relates to a cryopreservation solution for induced pluripotent stem cells, its application and cryopreservation method; CN201410676389.1 relates to a cryopreservation solution for human adipose-derived mesenchymal stem cells; CN201510443887.6 relates to a cryopreservation solution for stem cells and stem cell cryopreservation method; CN201510037248.X relates to a neural stem cell cryopreservation soluti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 滕理送金科涛蓝欢荣陶锋黄黎明吕杰青景元明王伟胡耕远
Owner SHAOXING PEOPLES HOSPITAL
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