Human bone marrow cell processing kit and cell processing method

Abstract

The invention relates to a human bone marrow cell processing kit and a cell processing method. The human bone marrow cell processing kit is characterized by consisting of the following three reagents: No.1 reagent which, as a thinner, is 0.5-20% of a sodium chloride injection or PBS (poly butylenes succinate) liquid, No.2 reagent which, as a precipitator, is 0.5-20% of hydroxyethyl starch or methylcellulose, and No.3 reagent which, as layering liquid, is prepared from ficoll and meglumine diatrizoate and is 1.0-1.2 in density. The cell processing method comprises the following steps: adding marrow containing sodium citrate anti-coagulation liquid to a high-capacity nutrient solution bottle which contains the No.1 reagent, and then adding the No.2 reagent and uniformly shaking; standing by, sucking upper cell liquid after layering and centrifuging for 1-20min, thinning the cell liquid by virtue of normal saline after the cell liquid is centrifuged and concentrated and paving the normal saline on the upper layer of No.3 liquid, then centrifuging so that a stem cell layer is separated, collecting the stem cell layer, and cleaning cells by virtue of normal saline for later use. The cell processing method is strong in operability, high in clinical safety and convenient for clinical popularization; and the kit is easy for storage and transportation, applicable to industrial production, and convenient and rapid to use.

Description

technical field [0001] The invention relates to biomedicine, in particular to a human bone marrow cell processing kit and a cell processing method. Background technique [0002] With the continuous development of modern science and technology, the separation technology for human blood cells has also been greatly developed. At present, there are mainly the following processing methods: immunomagnetic bead method, flow cytometry method, blood cell separation machine method and culture Amplification method, the above method has high cost and is not suitable for low- and middle-income patients. It will damage the activity of the cells themselves and affect the efficacy of cell therapy. The patient bears a certain risk of life. The pollution rate is high. long-term defects. [0003] CN1948467A discloses a kit for isolating bone marrow mononuclear cells. The disadvantage is that the number of stem cells in the blood is not enough for clinical treatment due to the use of this pate...

AI Technical Summary

Problems solved by technology

[0002] With the continuous development of modern science and technology, the separation technology for human blood cells has also been greatly developed. At present, there are mainly the following processing methods: immunomagnetic bead method, flow cytometry method, blood cell separation machine method and culture Amplification method, the above method has high cost and is not suitable for low- and middle-income patients. It will damage the activity of the cells themselves and affect the efficacy of cell therapy. The patient bears a certain risk of life. The pollution rate is high. long-term defects

[0003] CN1948467A discloses a kit for isolating bone marrow mononuclear cells. The disadvantage is that the number of stem cells in the blood is not enough for clinical treatment due to the use of this patent. Therefore, the patent has carried out cell culture, and the cell contamination rate of culture High, in vitro simulating the in vivo environment to make the cells expand, resulting in the cultured cells having a shape and no function, which cannot be used for clinical treatment; CN101144070A discloses a bone marrow umbilical cord blood stem cell in vitro separation kit and its application method, and its disadvantage is that the patent The lin antibody was added to the reagent, which greatly increased the cost. After it was put into clinical use, it brought a burden to low- and middle-income patients, resulting in patients who could not afford to see a doctor and receive cell therapy, and the use of lin antibody caused a large cost to cell cleaning. Burden, what will happen to this substance after it enters the human body is still unknown, and further research is needed

[0004] In summary, the existing cell separation technology mainly has the following problems: 1. The treatment cost is high

The final cell suspension is large in volume and poor in quality, and can only be used for autologous subcutaneous injection; 4. The time to obtain cells is too long

It can be used for clinical treatment after one week of culture and amplification. If the time is too long, it will delay the best time for treatment, affect the treatment effect, and increase the pollution rate during the culture period; 5. Traditional separation methods are limited to laboratory and scientific research use

Many separation methods are cumbersome to operate and have high professional requirements for personnel, which are time-consuming and laborious

Technical improvement is still needed from experimental methods to clinical applications; 6. Cannot be industrialized

The high cost of the experiment, the high probability of contamination in the operation process, and the unprepared storage and transportation conditions of the cell fluid lead to a limited amount of extracted cell fluid, which cannot meet the large demand for clinical treatment; The specificity of which blood to separate is not strong (because there are certain differences between human blood and animal blood cells), and the raw materials use biological products whose quality is difficult to control, etc.