Three-dimensional dynamic co-culture system for amplifying hematopoietic stem cells

Abstract

The invention provides a three-dimensional dynamic co-culture system for amplifying hematopoietic stem cells. Microcapsules are used as a growth system of trophoblast cells, and hematopoietic stem cells and the trophoblast cells are cultured in a three-dimensional dynamic condition. The dynamic environment is beneficial to transmission of nutrient substances, and proliferation of the hematopoieticstem cells is promoted. The three-dimensional co-culture environment is closer to the in-vivo hematopoietic microenvironment, and the nourishing and supporting effect of the trophoblast cells to thehematopoietic stem cells is closer to an in-vivo mode, so that proliferation of the hematopoietic stem cells and maintenance of cell activity are facilitated. Meanwhile, semi-permeable membranes of the microcapsules can effectively isolate immune rejection of the trophoblast cells and the hematopoietic stem cells and effectively isolate the trophoblast cells and the hematopoietic stem cells, and separation and harvest of the hematopoietic stem cells after the culture is finished are facilitated. The method for three-dimensional culture and expansion of hematopoietic stem cells provided by theinvention is expected to solve the technical bottleneck of insufficient number of hematopoietic stem cells during clinical use.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for expanding hematopoietic stem cells. Background technique [0002] Hematopoietic stem cells are the earliest stem cells used in clinical treatment. The CD34 markers are specifically expressed on their surface. They are used to treat malignant blood diseases, immunodeficiency syndrome, thalassemia, severe trauma and other diseases, and have achieved good therapeutic effects. . Hematopoietic stem cells come from a wide range of sources and can be obtained from bone marrow, peripheral blood, and umbilical cord, but their absolute number is relatively small. For example, the hematopoietic stem cells from a baby's umbilical cord blood can usually only satisfy children and low-weight adult patients, and it is difficult to meet the needs of adult transplantation. [0003] In order to expand the scope of hematopoietic stem cell transplantation, in vitro expansion of hematopoieti...

AI Technical Summary

Problems solved by technology

At present, the conventional methods for in vitro expansion of hematopoietic stem cells are mostly two-dimensional culture, using high-dose multi-cytokines joint culture, or using trophoblast cells (including mesenchymal stem cells, AGM cells, etc.) and multi-cytokines joint culture, these The method has obvious limitations: under the two-dimensional culture method, the cells lose their in vivo characteristics, and to a certain extent, it is easy to cause the differentiation of hematopoietic stem cells; there will be rejection between trophoblast cells and hematopoietic stem cells from different sources, which will affect the cell viability; Hematopoietic stem cells will adhere to the trophoblast cells, which is not conducive to the separation and harvest of the two types of cells; under static co-culture conditions, the culture medium lacks effective mixing, resulting in uneven nutritional components and metabolites

Some researchers have used microcarriers and calcium alginate microgel beads as three-dimensional culture substrates, but microcarriers still cannot avoid the direct contact between cells, resulting in difficulties in later separation, and the large number of micropores in calcium alginate microgel beads cannot prevent the culture process. Cell shedding into the culture medium and direct contact with hematopoietic stem cells

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