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Reagent box used for preparing bone marrow interstitial stem cell

A kit and stem cell technology, applied in bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve problems such as slow progress in scientific research

Active Publication Date: 2007-04-18
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 no kit for preparing bone marrow mesenchymal stem cells, which makes the progress of related research work relatively slow

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1. Preparation of a kit for preparing bone marrow mesenchymal stem cells and detection of its separation effect

[0018] 1. Preparation of kits for preparing bone marrow mesenchymal stem cells

[0019] 1. Solution preparation

[0020] The kit for preparing bone marrow mesenchymal stem cells of the present invention includes the following reagents:

[0021] (1) Diluent and washing liquid (abbreviated as dilution / washing liquid) are both: normal saline with a mass / volume percentage concentration of 0.9%;

[0022] (2) Separation liquid A is: the mass / volume percentage concentration is the 6.0% hydroxyethyl starch solution;

[0023] (3) Separation solution B is: Percoll solution with a density of 1.073g / mL. The preparation method is: take 9 parts of Percoll stock solution under sterile conditions, add 1 part of 1.5mmol / L PBS, mix well, and then take Wherein 0.56 part, add 0.44 part of 0.15mmol / L PBS, fully mix, obtain the Percoll solution that density is 1.073g / m...

Embodiment 2

[0047] Example 2. Preparation of a kit for preparing bone marrow mesenchymal stem cells and detection of its separation effect

[0048] 1. Preparation of kits for preparing bone marrow mesenchymal stem cells

[0049] 1. Solution preparation

[0050] The kit for preparing bone marrow mesenchymal stem cells of the present invention includes the following reagents:

[0051] (1) Diluent and washing liquid (referred to as dilution / washing liquid) are both: PBS, the formula is NaCl 0.8 g, KCl 0.2 g, NaCl 2 HPO 4 12H 2 O 2.9 g, KH 2 PO 4 0.2 g, add deionized water to make up to 1000 mL, autoclave;

[0052] (2) Separation solution A: a methylcellulose solution with a mass / volume concentration of 0.5%, the formula is: adding 0.5 g of methylcellulose into 100 mL of 0.9% mass / volume concentration of normal saline, and autoclaving After that, place it at 4°C and dissolve it;

[0053] (3) Separation solution B: polysucrose-sodium diatrizoate (ficoll-hypaque) solution (trade name F...

Embodiment 3

[0060] Example 3. Preparation of kit for preparing bone marrow mesenchymal stem cells and detection of its separation effect

[0061] 1. Preparation of kits for preparing bone marrow mesenchymal stem cells

[0062] 1. Solution preparation

[0063] The kit for preparing bone marrow mesenchymal stem cells of the present invention includes the following reagents:

[0064] (1) Diluent and washing liquid (referred to as dilution / washing liquid) are: Hank's and other solutions, the formula is: NaCl8g, KCl 0.4g, NaCl 2 HPO 4 ·H 2 O 0.06g, KH 2 PO 4 0.06g, NaHCO 3 0.35g, add deionized water to make up to 1000mL, after autoclaving, store at 4°C;

[0065] (2) separating liquid A: the mass / volume percentage concentration is the methyl cellulose solution of 0.55%;

[0066] (3) Separation solution B: polysucrose-sodium diatrizoate solution with a density of 1.077g / mL;

[0067] (4) Cell culture medium: cell culture medium A: low-sugar DMEM liquid medium; cell culture medium B: ne...

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Abstract

This invention discloses a kit used for preparing marrow desmohemoblast stem cell. This kit includes dilution, eluant, separating medium which consist of separating medium A and separating medium B, cell culture fluid A, cell culture fluid B and cell dissociation buffer A; 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 Percoll liquid whose density is 1.073g / mL or ficoll- hypaque solution whose density is 1.077g / mL; cell culture fluid A is low carbohydrates DMEM bouillon or DF12 nutritive medium; cell culture fluid B is fetal calf serum, new-born calf serum or calf serum; cell dissociation buffer A is parenzyme liquid whose quality / volume percentage concentration is 0.4-0.6%.

Description

technical field [0001] The invention relates to a kit, in particular to a kit for preparing bone marrow mesenchymal stem cells. Background technique [0002] For a long time, cardiovascular disease has been one of the main causes of human death, among which the death caused by acute myocardial infarction and chronic heart failure accounted for more than 50% of cardiovascular disease mortality. Due to the lack of regeneration ability of mature cardiomyocytes, after myocardial infarction, the number of cardiomyocytes decreases, and the infarcted area is replaced by scar tissue with non-contractile function through fibrous tissue proliferation, resulting in degenerative left ventricular remodeling and decreased cardiac function. , eventually leading to congestive heart failure. At present, the commonly used clinical treatment methods for acute myocardial infarction and chronic cardiac insufficiency after myocardial infarction mainly include drug therapy, medical interventional...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/08C12N5/0775
Inventor 裴雪涛王韫芳闫舫南雪白慈贤施双双管利东
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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