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Human marrow, cord blood or peripheral blood stem cells treating kit and stem cells separating method

A peripheral blood stem cell and umbilical cord blood technology, which is applied to blood/immune system cells, bone/connective tissue cells, animal cells, etc. The effect of low cost and strong operability

Inactive Publication Date: 2011-08-17
唐明淇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0033] The object of the present invention is to provide a kind of human bone marrow, umbilical cord blood or peripheral blood stem cell separation kit and the method for using the kit to separate human bone marrow stem cells, umbilical cord blood stem cells or peripheral blood stem cells, to improve human bone marrow, umbilical cord blood or peripheral blood The isolation quantity and recovery rate of stem cells, while overcoming the high cost of the existing technology, the presence of markers, the need to mobilize agents to cause pain to patients, long acquisition time, cumbersome and unsuitable for clinical use, etc.

Method used

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  • Human marrow, cord blood or peripheral blood stem cells treating kit and stem cells separating method
  • Human marrow, cord blood or peripheral blood stem cells treating kit and stem cells separating method
  • Human marrow, cord blood or peripheral blood stem cells treating kit and stem cells separating method

Examples

Experimental program
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Effect test

Embodiment 1

[0069] Isolation of Human Cord Blood:

[0070] Kit composition:

[0071] No. 1 solution: Diluent: PBS solution: 4g sodium chloride, 0.1g potassium chloride, 1.445g disodium hydrogen phosphate 12 water, 0.1g potassium dihydrogen phosphate Dissolve the above reagents with a small amount of water, then add water to 500ml, that is obtained, adjust the pH to 7-7.5 with 5.6% sodium carbonate before use. Add 3% cerebroprotein hydrolyzate (commercially available, prepared as 3% aqueous solution).

[0072] No. 2 solution: Precipitating agent: 6% hydroxyethyl starch (commercially available)

[0073] No. 3 liquid: Separation liquid: make polysucrose and diatrizoate into a separation liquid with a density of 1.075 g / ml.

[0074] The above-mentioned No. 1 and No. 2 liquids were sterilized at 100°C-130°C for 10 minutes-50 minutes, and the endotoxin content was detected to be ≤0.5EU / ml before being bottled. The above-mentioned No. 3 liquid was sterilized at 100°C-130°C for 10 minutes-50 ...

Embodiment 2

[0082] To isolate human bone marrow:

[0083] Kit composition:

[0084] No. 1 solution: Diluent: PBS solution: 4g sodium chloride, 0.1g potassium chloride, 1.445g disodium hydrogen phosphate 12 water, 0.1g potassium dihydrogen phosphate Dissolve the above reagents with a small amount of water, then add water to 500ml, that is To obtain it, adjust the pH to 7-7.5 with 5.6% sodium carbonate before use, and add 3% cerebroprotein hydrolyzate (commercially available, formulated as a 3% aqueous solution).

[0085] No. 2 solution: Precipitating agent: 6% hydroxyethyl starch (commercially available)

[0086] No. 3 liquid: Separation liquid: make polysucrose and diatrizoate into a separation liquid with a density of 1.075 g / ml.

[0087] The above-mentioned No. 1 and No. 2 liquids were sterilized at 100°C-130°C for 10 minutes-50 minutes, and the endotoxin content was detected to be ≤0.5EU / ml before being bottled. The above-mentioned No. 3 liquid was sterilized at 100°C-130°C for 10 m...

Embodiment 3

[0094] Isolation of Human Peripheral Blood:

[0095] Kit composition:

[0096] No. 1 solution: Diluent: PBS solution: 4g sodium chloride, 0.1g potassium chloride, 1.445g disodium hydrogen phosphate 12 water, 0.1g potassium dihydrogen phosphate Dissolve the above reagents with a small amount of water, then add water to 500ml, that is To obtain it, adjust the pH to 7-7.5 with 5.6% sodium carbonate before use, and add 3% cerebroprotein hydrolyzate (commercially available, formulated as a 3% aqueous solution).

[0097] No. 2 solution: Precipitating agent: 6% hydroxyethyl starch (commercially available)

[0098] No. 3 liquid: Separation liquid: make polysucrose and diatrizoate into a separation liquid with a density of 1.075 g / ml.

[0099] The above-mentioned No. 1 and No. 2 liquids were sterilized at 100°C-130°C for 10 minutes-50 minutes, and the endotoxin content was detected to be ≤0.5EU / ml before being bottled. The above-mentioned No. 3 liquid was sterilized at 100°C-130°C f...

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Abstract

The invention relates to a human marrow, cord blood or peripheral blood stem cell treating kit and a stem cell separating method. The kit consists of liquid 1, liquid 2 and liquid 3. Based on the prior art, brain protein hydrolysate is added into the liquid 1, so that the stem cells recovery quantity of the treating sample is effectively improved. The kit is used for separating the human marrow from the cord blood or the peripheral blood. Aiming at a cord blood sample, the separated cell recovery sum is improved by 5.9-15.8%, and the separated CD34 stem cell recovery sum is improved by 5.0-8.8%; aiming at a marrow sample, the separated cell recovery sum is improved by 8.7-21%, and the separated CD34 stem cell recovery sum is improved by 8.4-11%; and aiming at a peripheral blood sample, the separated cell recovery sum is improved by 23.1-46.7%, and the separated CD34 stem cells recovery sum is improved by 10.8-25.6%. Due to the improvement of the target stem cells separating and recovering ratio, the human marrow samples, cord blood samples or peripheral blood stem cell samples can be effectively saved, and the use efficiency of the samples can be improved.

Description

technical field [0001] The invention relates to a kit for separating stem cells in human bone marrow, umbilical cord blood or peripheral blood in vitro and a method for separating stem cells, belonging to the field of biomedical technology applications. Background technique [0002] In the prior art, methods for separating human blood cells include immunomagnetic bead method, flow cytometry method, blood cell separator method, culture expansion method and other methods. [0003] immunomagnetic bead method [0004] Known antibodies are coated on magnetic bead microparticles, and the magnetic bead microparticles are mixed with human blood. Antigen-antibody-positive cells adhere to the magnetic beads, pass through a magnetic tube, and the magnetic beads are adsorbed on the tube wall; after the other cells that are not bound to the magnetic beads flow away, remove the magnetic properties of the tube, and collect all the cells containing the magnetic beads . Problems: high cos...

Claims

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Application Information

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IPC IPC(8): C12N5/0789C12N5/0775
Inventor 唐明淇
Owner 唐明淇
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