Human bone marrow and umbilical cord blood stem cell treatment kit and stem cell separation method

Abstract

The invention discloses a human bone marrow and umbilical cord blood stem cell treatment kit. The stem cell treatment kit is composed of a No.1 liquid and a No.2 liquid, the percentage (%) is weight percentage (W / W), the density unit is g / ml, the solvent of the No.1 liquid and the solvent of the No.2 liquid are water; the No.1 liquid consists of 0.1%-30% of hydroxyethyl starch or 0.1%-30% of methyl cellulose; the No.2 liquid consists of a layering liquid which is prepared from polysucrose and meglumine diatrizoate, and has a density of 1.0-1.2 g / ml. In the implementation process, the reaction time can be shortened, reagents used in the scheme are fewer, and the number of obtained stem cells is larger.

Description

technical field [0001] The invention belongs to the technical field of biomedical technology application, and specifically relates to a human bone marrow and umbilical cord blood stem cell processing kit and a stem cell separation method. 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 cost, not suitable...

AI Technical Summary

Problems solved by technology

Existing problems: high cost, not suitable for low- and middle-income patients

Existing problems: mobilization agents increase the burden of treatment, the volume of cell fluid after separation is too large, and the patient bears a certain risk of life

Existing problems: high pollution rate, stem cells differentiate into unknown stem cells in an unknown direction, and it takes a long time

Disadvantages of this method of separating cells: the above method needs to be adjusted for chicken, cow, and human blood, and the accuracy is worrying. Due to the addition of surfactants, the isolated cells are unknown and can only be used for simple cell experiments.

Disadvantages of the stem cell separation liquid and stem cell separation method: working liquid needs to be prepared, density needs to be adjusted, and only cells with a density within the range of 1.083 g / ml can be collected, that is, the collected cells are more complicated and can be used for treatment. The number of target cells is small, and for umbilical cord blood, using this method to remove red blood cells is not clean, and the final clinical application will cause rejection in patients

[0015] Application number: 200610114475.9 Name: Kit for isolating bone marrow mononuclear cells: Disadvantages of using this kit: using this kit to directly separate stem cells in blood, the amount is not enough for clinical treatment, so this method is used for cell culture, The contamination rate of the cultured cells is high, and the in vitro simulation of the in vivo environment causes the cells to proliferate, resulting in the cultured cells having a shape and no function, which cannot be used for clinical treatment

Disadvantages: The HISTOPAGUE 1077 used in this kit and its application method is a lymphocyte separation medium with a density of 1.077g / ml. Most of the cells after this separation are lymphocytes, which leads to a small number of stem cells in the cells that can be used for treatment , affecting clinical efficacy

Disadvantages: The use of the kit adds lin antibody, which leads to a great increase in cost. After it is put into clinical use, it will cause economic burden to low- and middle-income patients, causing patients to be unable to see a doctor and accept cell therapy, and the use of lin antibody to treat Cell cleaning has caused a large burden, and it is still unknown what will happen to this substance after it enters the human body

[0032] The kits or methods disclosed in the above relevant patent documents have disadvantages such as complex sources of raw materials, cumbersome preparation, lack of specificity for the separation of blood (because there are certain differences between human blood and animal blood cells), and difficulty in controlling the use and treatment of the obtained separation liquid.

[0033] In the Chinese invention patent application with the application number CN201110026680.0, a human bone marrow and umbilical cord blood stem cell processing kit and stem cell separation method are disclosed. longer time

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