Culture medium and culture method for induced differentiation of CD34+ umbilical cord blood mononuclear cells

Abstract

The invention discloses a culture medium for induced differentiation of CD34+ umbilical cord blood mononuclear cells. The culture medium consists of a basal culture medium and a composite additive; the composite additive consists of following components: 40-120 ng / ml of SCF; 4-8 ng / ml of IL3; 3-5 IU of Epo; 1-2 [mu]M of hydrocortisone or dexamethasone; 30-35 ng / ml of Flt3L; and 30-40 ng / ml of IGF-1. The invention further discloses a culture method for induced differentiation of the CD34+ umbilical cord blood mononuclear cells. The culture method comprises the following steps: (1) inoculating CD34+ umbilical cord blood mononuclear cells into a first-stage culture medium for culture; (2) from the seventh day, adding a second-stage culture medium for culture; (3) from the eleventh day, addinga third-stage culture medium for culture; and performing culture to the eighteenth day to obtain erythroid specific cells. According to the culture medium and the culture method, the induced differentiation efficiency of the CD34+ umbilical cord blood mononuclear cells can be remarkably improved, the differentiation and amplification time of the CD34+ umbilical cord blood mononuclear cells is shortened, and the erythroid specific cells are rapidly and efficiently obtained.

Description

technical field [0001] The invention relates to a culture medium and a culture method for inducing differentiation of CD34+ umbilical cord blood mononuclear cells, belonging to the technical field of cell culture. Background technique [0002] Since the world's first successful umbilical cord blood stem cell transplantation in 1989, more than 1,000 cases of umbilical cord blood hematopoietic stem cell transplantation have been used to treat leukemia and other diseases. Compared with bone marrow and peripheral blood, umbilical cord blood has many advantages as a source of transplantation: first, the ratio of primitive stem / progenitor cells in umbilical cord blood is higher than that of bone marrow and mobilized peripheral blood; secondly, the transmission of viral diseases in transplantation is reduced, and the The incidence of graft-versus-host disease (GVHD) has been reduced; in addition, cord blood has a wide range of sources and is convenient to collect, so research on co...

AI Technical Summary

Problems solved by technology

However, due to the small amount of a single cord blood, it is not enough to meet the needs of adult transplantation, so people hope to increase the number of hematopoietic stem / progenitor cells in cord blood through in vitro expansion

[0003] In the in vitro expansion of umbilical cord blood hematopoietic stem / progenitor cells, CD34+ enriched cells can be used as the starting cells for culture, but the culture medium used in the conventional in vitro culture of CD34+ enriched cells is not ideal, and the cell proliferation rate is slow , low expression, limited number of proliferating cells, long induction differentiation time, low efficiency and other technical defects

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