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Bone marrow cell culture medium

A bone marrow cell and culture medium technology, which is applied in the field of bone marrow cell culture medium formula, can solve the problems of poor dispersion, high cost, and low bone marrow chromosome division index, and achieve a clear background, many cleavage phases, good shape and dispersion Effect

Active Publication Date: 2014-03-26
南昌艾迪康医学检验实验室有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the interference of a large number of fat particles in the bone marrow, the cell cycle of various cell lines in the bone marrow is not fixed, not uniform, and it is difficult to treat them differently, resulting in low chromosome division index, short and thick chromosomes, poor dispersion, and cost higher

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of bone marrow cell culture medium

[0026] Penicillin / streptomycin, fetal bovine serum, and human lymphoma cell cultures were added to the basal medium. Wherein the basal medium is RPMI1640 medium, and the consumption of each added component is as follows:

[0027] Penicillin / Streptomycin 5-15ul / ml

[0028] Fetal bovine serum 60-140 ul / ml

[0029] Human Lymphoma Cell Culture 60-140 ul / ml

[0030] Wherein, penicillin can be selected from 10000 U / ml, and streptomycin can be selected from 10000 μg / ml. In other embodiments, other concentrations of penicillin and streptomycin can be selected for the formulation of the medium.

[0031] Wherein, the human lymphoma cell culture is prepared according to the following method: the cells are resuscitated and subcultured, and when the culture medium of the subcultured cells turns yellow, the cells are collected and centrifuged, and after centrifugation, the supernatant is collected and filtered to obtain t...

Embodiment 2

[0043] Example 2 Bone marrow cell culture and chromosome preparation

[0044] In this example, the bone marrow cells were cultured according to the following method, and after harvesting, they were used for chromosome G-band preparation. In other embodiments, other methods of culturing bone marrow cells and chromosome preparations can also be used. The method described in this embodiment is:

[0045] (1) Preparation of bone marrow culture medium: preparation according to the method of Example 1;

[0046] (2) Inoculation: inoculate the bone marrow cells into the medium described in step 1;

[0047] (3) Termination of culture: the cells were terminated by colcemid;

[0048] (4) Collection of bone marrow cell culture;

[0049] (5) Chromosomal specimen preparation: Chromosomal specimens with G-banding were obtained after staining with dyes.

[0050] Among them, the bone marrow cells were divided into 1~3×10 6 Cells / ml density was inoculated into bone marrow medium, placed ...

Embodiment 3

[0068] Embodiment 3 Comparative Experiment 1

[0069] According to the method of Example 1, three groups of culture media were prepared, namely culture medium 1, culture medium 2, and culture medium 3, and a control group was set up for bone marrow culture. The culture medium (medium 4) of the control group did not contain human Lymphoma cell culture. Bone marrow cells from the same sample were taken and cultured in medium 1 to medium 4 according to the method in Example 2, and the chromosome G band was produced according to the method in Example 2, and the results were observed under a microscope. The microscope model used is: Leica DM2500.

[0070] in:

[0071] The formulation of medium 1 is:

[0072] Basal medium: RPMI1640 medium

[0073] Penicillin / Streptomycin 5ul / ml

[0074] Fetal bovine serum 60ul / ml

[0075] Human lymphoma cell culture 60 ul / ml

[0076] The formulation of medium 2 is:

[0077] Basal medium: RPMI1640 medium

[0078] Penicillin / Streptomycin ...

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Abstract

The invention discloses a bone marrow cell culture medium formula for karyotype analysis in the field of medical examination. According to the invention, penicillin / streptomycin, fetal calf serum and human lymphoma cell culture are added into a basal culture medium. Chromosome G-band produced by a bone marrow cell prepared by utilizing the culture medium disclosed by the invention has clearer background, more mitotic phases and more excellent forms and dispersity and facilitates analysis. For bone marrow karyotype analysis, the culture medium formula for karyotype analysis has the time-saving and labor-saving advantages, higher accuracy and broader application prospect in the field of medical examination.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a bone marrow cell culture medium formula for chromosome karyotype analysis in the field of medical testing. Background technique [0002] G-banding is called G-banding technology because the chromosomes are mainly stained with Giemsa dye, and the banding patterns displayed are distributed on the entire chromosome. People use various methods and different dyes to process chromosome samples to make each chromosome appear light and dark, or the technology of different shades of banding is called banding technique (banding technique). Since the 1970s, the banding technology has been greatly developed, and among the many banding technologies (Q band, G band, C band, R band, T band), the G band is currently widely used. belt type. [0003] Studies have found that human chromosome specimens are treated with reagents such as trypsin, Na0H, citrate or urea, and then stained with Giemsa, wh...

Claims

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Application Information

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IPC IPC(8): C12N5/077G01N1/28
Inventor 程建兵夏成青陈红梅郭福晓生帅
Owner 南昌艾迪康医学检验实验室有限公司
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