Bone marrow cell culture medium

A bone marrow cell and culture medium technology, applied in the direction of cell culture active agent, bone/connective tissue cells, culture process, etc., can solve the problems of poor dispersion, high cost, low bone marrow chromosome division index, etc., to achieve clear background, split The effect of multiple phases, good shape and dispersion

Inactive Publication Date: 2016-05-25
谷超
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  • Summary
  • 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 and uniform, and it is difficult to treat them differently, resulting in low chromosome division index, short and thick chromosomes, poor dispersion, and low cost. higher

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of bone marrow cell culture medium

[0021] The basal medium is RPMI1640 medium containing fetal bovine serum, human lymphoma cell culture and compound CephaloziellinN; the concentration of fetal bovine serum in the basal medium is 100 μl / ml, and the concentration of compound CephaloziellinN in the basal medium is 25 μg / ml ml. It also contains penicillin at a concentration of 5000U / ml. Penicillin can be replaced by streptomycin at a concentration of 6000 μg / ml. The concentration of human lymphoma cell culture in the basal medium is 120 μl / ml. The preparation method of human lymphoma cell culture is as follows: recover and subculture human lymphoma cells, and collect the cells when the color of the subcultured cell culture medium turns yellow And centrifuge, after centrifugation, collect the supernatant. More detailed preparation steps are as follows:

[0022] Step S1, clean the ultra-clean bench with Promethieme, and irradiate it with ultravio...

Embodiment 2

[0031] Embodiment 2: the contrast of embodiment 1, do not add CephaloziellinN

[0032] The basal medium is RPMI1640 medium, containing fetal bovine serum and human lymphoma cell culture; the concentration of fetal bovine serum in the basal medium is 100 μl / ml. It also contains penicillin at a concentration of 5000U / ml. Penicillin can be replaced by streptomycin at a concentration of 6000 μg / ml. The concentration of human lymphoma cell culture in the basal medium is 120 μl / ml. The preparation method of human lymphoma cell culture is as follows: recover and subculture human lymphoma cells, and collect the cells when the color of the subcultured cell culture medium turns yellow And centrifuge, after centrifugation, collect the supernatant. More detailed preparation steps are as follows:

[0033] Step S1, clean the ultra-clean bench with Promethieme, and irradiate it with ultraviolet light for 30 minutes;

[0034] Step S2, take out the cryopreservation tube from the liquid nit...

Embodiment 3

[0042] Embodiment 3: effect embodiment

[0043] Use the bone marrow cell culture medium of embodiment 1 and 2 respectively to cultivate bone marrow cells according to the following method, and use for chromosome G-band preparation after harvesting:

[0044] (1) Prepare bone marrow culture medium: prepare according to the method of embodiment 1 and 2;

[0045] (2) Inoculation: inoculate bone marrow cells into the culture medium described in step 1;

[0046] (3) Termination of culture: the cells were terminated by colcemid;

[0047] (4) receiving bone marrow cell culture;

[0048] (5) Chromosomal specimen preparation: Chromosomal specimens with G-banding were obtained after staining with dyes.

[0049] Among them, the bone marrow cells were divided into 1~3×10 6 Inoculate into the bone marrow culture medium at a density of 1 / ml, put in 37 ℃, 5% CO 2 Incubator for 24 hours. The resulting bone marrow culture can be used for bone marrow chromosome preparation.

[0050] Where...

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Abstract

The invention discloses a bone marrow cell culture medium which comprises a basal culture medium containing fetal calf serum, human lymphoma cell culture and a compound Cephaloziellin N. The invention also provides a method for preparing bone marrow cells used in bone marrow chromosome sectioning, comprising the steps of: inoculating bone marrow cells with density of 1-3*106 / ml into the bone marrow cell culture medium, and putting into an incubator of 37 DEG C with 5% CO2 for culturing for 20-28 hours. When the bone marrow cells cultured by the bone marrow cell culture medium provided by the invention are used in chromosome G band sectioning, a clearer background, more division phases and better form and dispersion degree are obtained.

Description

technical field [0001] The invention belongs to the field of cell culture, in particular to a bone marrow cell culture medium. Background technique [0002] In recent years, with the development of molecular biology and cytogenetics, bone marrow karyotype analysis has played an increasingly important role in the diagnosis, treatment and prognosis of blood system diseases. 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 and uniform, and it is difficult to treat them differently, resulting in low chromosome division index, short and thick chromosomes, poor dispersion, and low cost. higher. [0003] Therefore, it is particularly important to establish a bone marrow culture medium formula with clearer background, more cleavage phases, good shape and dispersion. Contents of the invention [0004] The purpose of the present invention is to overcome the deficiencies of the prior ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0669C12N2500/84C12N2501/999
Inventor 谷超
Owner 谷超
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