A kind of human primary cell culture medium and its application

Abstract

Provided are a primary cell culture medium, use thereof for culturing a human primary cell in vitro, and method for utilizing the culture medium to culture a human primary B-cell acute lymphoblastic leukemia (B-ALL) cell in vitro. The primary cell culture medium comprises a base culture medium and cell factors: human FLT3L, human IGFl, human IL-7 and human IL-6, and is serum-free.

Description

technical field [0001] The present invention relates to the technical field of cell culture, in particular to a primary cell culture medium, the use of the medium for culturing human primary cells in vitro, and the use of the culture medium for culturing primary human acute lymphoblastic leukemia (B- ALL) cell method. Background technique [0002] B-cell acute lymphoblastic leukemia (B-ALL, B-cell acute lymphoblastic leukemia), also known as precursor B-cell (pre B-cell) acute lymphoblastic leukemia, is a malignant tumor derived from B-cell progenitor cells. B-ALL is mainly a high-incidence cancer in children, and its incidence rate decreases in adults. Prognosis is good in juvenile patients with B-ALL, and the long-term survival rate (EFS, event-free survival) can reach 90%, but it is poor prognosis and low survival rate in B-ALL adults. In addition, in adult B-ALL patients, the effect of traditional chemotherapy is poor, and the mortality rate is about 60%. Therefore, t...

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Problems solved by technology

[0008] An article titled "Long–term culture of primary human lymphoblasticleukemia cells in the absence of serum or hematopoietic growth factors" in 2009 mentioned the in vitro culture medium of primary lymphocytic leukemia cells, which was supplemented with the cytokine IL -3. Serum-free IMDM medium of IL-7 and SCF; although this medium does not contain serum, however, the inventors of the present invention used this medium to carry out repeated experiments and found that the culture effect described in the article could not be achieved , that is, its stability needs to be investigated;

[0010] It should be noted that although serum is necessary for the survival and culture of traditional cells in vitro, when human primary cells are cultured in vitro, serum can accelerate the differentiation and aging of primary cells, especially hematopoietic stem cells and progenitor cells; Moreover, as far as the serum itself is concerned, even if it is the serum of the same brand, there may be obvious differences between different batches of serum, which makes the culture effect also have obvious differences, so the use of serum-containing medium has unclear medium components. The problem is that the practicability is relatively insufficient

[0011] However, co-cultivation with stromal cells is not only cumbersome, but also the growth rate of stromal cells (especially immortalized cell lines) far exceeds that of primary cells during the co-cultivation process, which competes with primary cells for nutrients in the medium. , thereby affecting the growth of primary cells; in addition, stromal cells activate the survival and expansion of primary cells in vitro through protein secretion or receptor binding, but these activation factors are uncontrollable with the cell state, differentiation, aging, and passage times of stromal cells. Factors are related, resulting in obvious differences in the culture effect of primary cells cultured by the stromal cell co-culture method, which is difficult to repeat and is not conducive to popularization and application

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