Reagent box used for separating bone marrow single nuclear cell

A kit and a single technology, applied in the field of kits for isolating bone marrow mononuclear cells, can solve problems such as slow progress in scientific research

Active Publication Date: 2010-10-06
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no kit for isolating bone marrow mononuclear cells, making the progress of related research work relatively slow

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1. Preparation of a kit for isolating bone marrow mononuclear cells and detection of its separation effect

[0017] 1. Preparation of a kit for isolating bone marrow mononuclear cells

[0018] 1. Solution preparation

[0019] The kit for isolating bone marrow mononuclear cells of the present invention includes the following reagents:

[0020] (1) Both the diluent and the washing liquid are: normal saline with a mass / volume percentage concentration of 0.9%;

[0021] (2) Separation liquid A is: the mass / volume percentage concentration is the 6.0% hydroxyethyl starch solution;

[0022] (3) Separation solution B is: polysucrose-sodium diatrizoate (ficoll-hypaque) solution (trade name Ficoll solution) with a specific gravity of 1.077g / mL;

[0023] (4) Restoration combination solution: restoration solution A: high-sugar DMEM medium, restoration solution B: fetal bovine serum, restoration solution C: 200mM L-glutamine; After the solution A and the repair solution B...

Embodiment 2

[0042] Example 2. Preparation of a kit for isolating bone marrow mononuclear cells and detection of its separation effect

[0043] 1. Preparation of a kit for isolating bone marrow mononuclear cells

[0044] 1. Solution preparation

[0045] The kit for isolating bone marrow mononuclear cells of the present invention includes the following reagents:

[0046] (1) Diluent and washing liquid are both: PBS, the formula is: NaCl 0.8 g, KCl 0.2 g, NaCl 2 HPO 4 12H 2 O2.9 g, KH 2 PO 4 0.2 g, add deionized water to make up to 1000 mL, autoclave;

[0047] (2) Separation solution A: a methylcellulose solution with a mass / volume percentage concentration of 0.5%. Preparation method: add 0.5g of methylcellulose into 100mL of normal saline, after autoclaving, place at 4°C, and wait until completely Just after dissolving;

[0048] (3) Separation solution B: Percoll solution with a specific gravity of 1.073 g / mL.

[0049] 2. Packing of each component

[0050] The specification of th...

Embodiment 3

[0059] Example 3. Preparation of a kit for isolating bone marrow mononuclear cells and detection of its separation effect

[0060] 1. Preparation of a kit for isolating bone marrow mononuclear cells

[0061] 1. Solution preparation

[0062] The kit for isolating bone marrow mononuclear cells of the present invention includes the following reagents:

[0063] (1) Diluent and washing liquid are both: Hank's solution, the preparation method is: NaCl 8g, KCl 0.4g, NaCl 2 HPO 4 .H 2 O0.06g, KH 2 PO 4 0.06g, NaHCO 3 0.35g, add deionized water to make up to 1000mL, after autoclaving, store at 4°C;

[0064] (2) separating liquid A is: the mass / volume percentage concentration is the 6.5% hydroxyethyl starch solution;

[0065] (3) Separation solution B is: polysucrose-sodium diatrizoate (ficoll-hypaque) solution (trade name Ficoll solution) with a specific gravity of 1.077g / mL;

[0066] (4) Repair solution: repair solution A: 1640 medium; the method of using the repair solutio...

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PUM

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Abstract

This invention discloses a kit used for separating marrow mononuclear cell. This kit includes dilution, eluant, separating medium which consist of separating medium A and separating medium B; the stated dilution is isotonic Na chloride, PBS or Hank's liquid; eluant is isotonic Na chloride, PBS or Hank's liquid; separating medium A is hydroxyethyl starch solution whose quality / volume percentage concentration is 5.5-6.5% or cellogran solution whose quality / volume percentage concentration is 0.45-0.55%; Separating medium B is ficoll- hypaque solution (trade name is ficoll solution) whose density is 1.077g / mL or Percoll liquid whose density is 1.073g / mL.

Description

technical field [0001] The invention relates to a kit, in particular to a kit for isolating bone marrow mononuclear cells. Background technique [0002] Limb ischemic diseases caused by arterial obstruction (arteriosclerotic obliterans of the lower extremities, diabetic limb ischemia, thromboangiitis obliterans, etc.) are common and refractory diseases in my country and even in the world. With the increase of the elderly population, the number of cases is gradually increasing. The risk factors associated with its pathogenesis include hypertension, hyperlipidemia, diabetes, smoking, lack of exercise, and population aging, etc. These risk factors are developing in a direction that is not conducive to disease control. According to reports from western countries, the incidence rate of this type of disease is as high as 15% among adults over 55 years old, and more than 1 / 3 of them have typical intermittent claudication symptoms; 12.1%. In the advanced stage of this disease, the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08C12N5/0786
Inventor 裴雪涛王韫芳南雪闫舫施双双管利东
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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