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Chromosome culture medium for quickly harvesting metaphase cells and application

A culture medium and chromosome technology, applied in the fields of cell biology and human medical genetics, can solve the problems of time-consuming, time-consuming, and uneven quality of diagnosis results, so as to facilitate karyotype analysis and clinical diagnosis, and shorten the time required for diagnosis. The effect of taking time and shortening the processing time

Inactive Publication Date: 2017-05-31
河北意和医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no uniform standard for chromosome culture media on the market, and the quality is uneven. Some culture media are of unstable quality, which affects the effect of chromosome preparation and analysis and diagnosis.
Chromosome examination is applicable to a wide range of people and requires a lot of examinations, but the preparation of chromosome specimens takes a long time, which not only affects the generation time of diagnostic results, but also affects the progress of other examinations for patients, and it is not convenient for major hospitals to carry out a large number of chromosome examinations. an examination
In the process of chromosome preparation, it is necessary to add colchicine to terminate the metaphase cells, and the processing time of colchicine is generally longer, which is a time-consuming step in the chromosome preparation process
Although there have been some reports of improved medium, but its application in chromosome preparation, the colchicine treatment time is still relatively long, generally 2-4 hours

Method used

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  • Chromosome culture medium for quickly harvesting metaphase cells and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 The preparation of a chromosomal medium for rapidly harvesting metaphase cells comprises the following steps:

[0022] (1) Prepare penicillin sodium and streptomycin sulfate according to the following method

[0023] Preparation of Penicillin Sodium Injection:

[0024] Penicillin Sodium for Injection (1.6 million units / bottle) 1

[0025] Sterile normal saline 4.5 ml

[0026] Streptomycin Sulfate Injection Preparation:

[0027] Streptomycin Sulfate for Injection (1 million units / bottle) 1

[0028] Sterile normal saline 5 ml

[0029] Use a disposable sterile syringe to draw sterilized saline, inject it into penicillin sodium or streptomycin sulfate bottle, shake it gently, and store it at -20°C after use;

[0030] (2) To prepare phytohemagglutinin (PHA) solution, draw a certain amount of sterilized distilled water with a disposable sterile syringe to dissolve 10 tubes (10 mg per tube) of PHA powder.

[0031] (3) Culture medium preparation

[0032] First, ...

Embodiment 2

[0033] Example 2 Culture and Chromosome Preparation of Human Peripheral Blood Lymphocytes (Application in Culture of Human Peripheral Blood Lymphocytes and Chromosome Karyotype Analysis).

[0034] (1) Peripheral blood collection: Disinfect the skin at the cubital vein with iodophor alcohol, and after it is completely dry, collect 2 ml of blood with a 2.5 ml disposable sterile syringe, draw 0.2 ml of heparin sodium injection immediately, put the syringe flat, and mix gently .

[0035] (2) Inoculation culture: After the medium returns to room temperature, remove the aluminum sheet on the medium with tweezers, disinfect the rubber stopper with alcohol, and place it until the alcohol is completely evaporated. Discard the first few drops of blood from the syringe, operate under an alcohol lamp, insert the needle into the medium bottle obliquely through the rubber stopper at an angle of 45 degrees, inject 0.6 ml of whole blood drop by drop, mix gently and set the temperature at 37°C...

Embodiment 3

[0046] Example 3 Comparison of lymphocyte transformation rate and lymphocyte division index in chromosomal medium

[0047] Two brands of chromosome culture medium commonly used by Hebei Provincial Children's Hospital and the First Hospital of Hebei Medical University were selected, numbered ET and YD. According to Example 1, the chromosome culture medium of the present invention was prepared, different batches of medium labels A1-A6 were selected, and 0.6 ml of the blood of the same individual was respectively inoculated, and cell culture and chromosome preparation were carried out according to Example 2. The comparison of cell culture effects is shown in Table 1. It can be seen from Table 1 that the transformation rate and division index of the chromosome culture medium of the present invention are better than those of the other two brands, and the growth and proliferation effect of the cells is better. And from the comparison between different batches of A1-A6, the numerica...

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Abstract

The invention discloses a chromosome culture medium for quickly harvesting metaphase cells. The chromosome culture medium for quickly harvesting the metaphase cells comprises the following active ingredients: an RPMI 1640 powder culture medium, calf serum, penicillin sodium, streptomycin sulphate, sodium bicarbonate and phytohemagglutinin(PHA), wherein the contents of various active ingredients are respectively as follows: 10.4 g / L of the RPMI 1640 powder culture medium, 100 ml / L of the calf serum, 400 to 600 mu l / L of the penicillin sodium, 400 to 600 mu l / L of the streptomycin sulphate, 2 g / L of the sodium bicarbonate and 100 to 120 mg / L of the phytohemagglutinin(PHA). The effects of different components and the PHA are fully played; a large amount of cells in a metaphase can be obtained; chromosome nuclear type analysis and clinical diagnosis are conveniently performed; meanwhile, the chromosome culture medium can support quickly and effectively harvesting the metaphase cells and shorten the processing time of colchicines, so that the time required for preparing chromosomes is shortened; help is provided for performing the chromosome nuclear type analysis and genetic diagnosis largely.

Description

technical field [0001] The invention relates to a chromosome culture medium, in particular to a chromosome culture medium for rapidly harvesting metaphase cells and an application thereof, belonging to the technical fields of cell biology and human medical genetics. Background technique [0002] Any abnormal number of chromosomes, even small structural aberrations, such as duplications, deletions, translocations, etc., will lead to the increase or absence of genes, resulting in corresponding clinical phenotypes. Such as multiple dysmorphic syndrome, symptoms include multiple deformities, mental retardation and abnormal growth and development, in addition there may be some changes in skin texture. Chromosomal aberrations can also lead to fetal stillbirth or miscarriage. Carriers of balanced chromosomal translocations with normal appearance may cause miscarriage, stillbirth, neonatal death or congenital malformation in their offspring. It has been found that more than 50% of...

Claims

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Application Information

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IPC IPC(8): C12N5/078
Inventor 王俊霞连伟光李丹丹张文昊祝岩波
Owner 河北意和医学检验实验室有限公司
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