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A kind of biological sample preservation solution and its preparation method and application

A biological sample and preservation solution technology, applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve the problems such as introduction of viruses and other microorganisms, maintenance of biological activity, difficulty in biological samples, etc., to meet long-term/long-distance The effect of transportation, increasing application prospects, and simple preparation method

Active Publication Date: 2022-07-01
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the isolation of MSCs needs to be carried out within a short time after the tissue leaves the mother, otherwise a sufficient amount of primary MSCs cannot be isolated, which will affect the subsequent culture and proliferation of MSCs
At present, physiological saline or cell culture medium is usually used as the preservation solution. The above-mentioned preservation solution can only provide certain nutrients or living environment in a short period of time, and long-term preservation cannot achieve effective protection. It is difficult for biological samples to maintain biological activity for a long time. , unable to isolate primary cells
In addition, the serum contained in the preservation solution can easily lead to the introduction of microorganisms such as viruses, which virtually increases the safety risk of the product

Method used

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  • A kind of biological sample preservation solution and its preparation method and application
  • A kind of biological sample preservation solution and its preparation method and application
  • A kind of biological sample preservation solution and its preparation method and application

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preparation example Construction

[0056] The present invention provides a method for preparing the above-mentioned biological sample preservation solution. The preparation method includes the following steps: stirring and mixing the high-glucose DMEM medium solution, compound electrolyte solution and heparin sodium, and then adding human serum albumin, ginseng Saponin, human umbilical cord mesenchymal stem cell culture supernatant, double antibody, stir and mix, and that's it.

[0057] The present invention also provides an application of the above-mentioned biological sample preservation solution in preparing and preserving biological sample reagents.

Embodiment 1

[0060] The preparation method of human umbilical cord mesenchymal stem cell culture supernatant is as follows:

[0061] Take a healthy umbilical cord, rinse it with normal saline until the surface is free of blood, remove the blood vessels, peel off the Wharton's glue, cut into pieces, culture by tissue block culture, at 37°C, 5% CO 2 Culture in the incubator until there are cells crawling out from around the tissue block, when the cells grow to 90%, digest with 0.25% trypsin, subculture to the 5th generation, and take the 5th generation of mesenchymal stem cells to expand and culture to a confluence rate. Supernatant at 95%, cell density 1.8 x 10 5 Collect the supernatant after the cells / mL, discard the precipitate, and filter with a 400-mesh filter to obtain the culture supernatant of human umbilical cord mesenchymal stem cells;

[0062] The preparation method of ginsenoside is as follows:

[0063] Take 500g of ginseng powder and add it to 12500mL of 70% ethanol solution. ...

Embodiment 2

[0066] The preparation method of human umbilical cord mesenchymal stem cell culture supernatant is as follows:

[0067] Take a healthy umbilical cord, rinse it with normal saline until the surface is free of blood, remove the blood vessels, peel off the Wharton's glue, cut into pieces, culture by tissue block culture, at 37°C, 5% CO 2 Culture in the incubator until there are cells crawling out from around the tissue block, when the cells grow to 95%, digest with 0.25% trypsin, subculture to the 5th generation, and take the 5th generation of mesenchymal stem cells to expand and culture to a confluence rate. 85% supernatant at a cell density of 2 x 10 5 Collect the supernatant after the cells / mL, discard the precipitate, and filter with a 400-mesh filter to obtain the culture supernatant of human umbilical cord mesenchymal stem cells;

[0068] The preparation method of ginsenoside is with embodiment 1;

[0069] 50 mL of high-glucose DMEM medium, 40 mL of compound electrolyte s...

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Abstract

The invention provides a biological sample preservation solution, which belongs to the technical field of biological sample preservation. Supernatant, double antibody. The preservation solution of the invention can ensure sufficient nutrient supply for tissue or blood cells, and prolong the in vitro preservation time of umbilical cord tissue, hair follicle tissue and menstrual blood samples. The preservation solution of the invention has a wide range of applications, and can make biological samples meet the requirements of long-term / long-distance transportation of biological samples under the condition of maintaining high survival rate and activity, thereby increasing the application prospect of mesenchymal stem cells.

Description

technical field [0001] The invention relates to the technical field of biological sample preservation, in particular to a biological sample preservation solution and a preparation method and application thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are an important member of the stem cell family, originating from the mesoderm in the early stage of development. More and more people's attention, is a widely used stem cells. [0003] Mesenchymal stem cells were initially found in the bone marrow, and subsequently found in many kinds of tissues during the occurrence and development of the human body, such as bone marrow, fat, synovium, bone, muscle, lung, liver, pancreas, umbilical cord, hair follicles and other tissues. After being isolated from tissues such as umbilical cord, mesenchymal stem cells have high proliferation ability and in vitro passage ability in vitro, which meets the requirement of a large number of MSCs for clinical treatment. [0004...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/021
Inventor 田娜张石林龚庆黄玉玲鞠帅
Owner JILIN TUO HUA BIOTECH
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