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Program induction method for promoting embryonic stem cell directional differentiation to hemopoietic stem cell

A technology of hematopoietic stem cells and embryonic stem cells, which is applied in the field of inducing embryonic stem cell-directed differentiation, can solve the problems of restricting the wide application of hematopoietic stem cell transplantation, limited sources of HLA-matched donors, and low differentiation efficiency of ES cells, so as to reduce social medical burden and save The effect of improving patient life and promoting functional development

Inactive Publication Date: 2005-09-28
SUN YAT SEN UNIV
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Problems solved by technology

Since allogeneic hematopoietic stem cell transplantation requires strict tissue matching, the source of HLA-matched donors is extremely limited, which limits the wide application of hematopoietic stem cell transplantation in clinical practice.
ES cells with developmental pluripotency can differentiate into hematopoietic stem cells in vitro, but ES cells have problems such as low differentiation efficiency and hematopoietic hypoplasia. It is urgent to invent a safe and effective method to induce cultured ES cells to direct towards hematopoietic stem cells Differentiate and promote its effective expansion and functional development, providing an ideal cell source for hematopoietic stem cell transplantation

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  • Program induction method for promoting embryonic stem cell directional differentiation to hemopoietic stem cell

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Embodiment Construction

[0008] The present invention will be further described below in conjunction with accompanying drawing.

[0009] Isolation of yolk sac (embryo age: mouse 8-11 days / human 4-6 weeks) and fetal liver (embryo age: mouse 10-14 days / human 8-12 weeks) from mouse embryos or human aborted embryos Tissues, adult mouse bilateral femurs or adult bone marrow tissue were extracted, yolk sac, fetal liver and bone marrow single cell suspensions were prepared by enzymatic digestion or physical separation, and mononuclear cells were separated by density gradient centrifugation with 15% Fetal bovine serum in DMEM at 37°C, 5% CO 2 1. Carry out stromal cell culture under saturated humidity conditions, change the medium once every 3-5 days, and cultivate to a sub-confluent state. Take stromal cells in good growth state to prepare trophoblast and serum-free conditioned medium. At the same time, the type of stromal cells and The composition ratio is identified.

[0010] The yolk sac, fetal liver and...

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Abstract

The programmed inducing process of promoting directional differentiation of embryo stem cell to hemopoietic stem cell includes taking tissue, culturing stromal cell, collecting no-serum condition culture liquid, program inducing the directional differentiation of embryo stem cell to hemopoietic stem cell, sorting hemopoietic stem cell and other steps. The process features that embryo stem cell is cultured to form embryo proper and hemopoietic stromal cells in different growth stage are separated and cultured from yolk sac, embryo liver and adult myeloid tissue to form the cell layer in sub-fusion state. The present invention makes it possible to induce producing great amount of completely grown hemopoietic stem cells for clinical application.

Description

[technical field] [0001] The invention relates to a method for inducing directional differentiation of embryonic stem cells (ES cells), which can improve the efficiency of directional differentiation of ES cells to hematopoietic stem cells and promote the functional development of ES-derived hematopoietic stem cells. [Background technique] [0002] Hematopoietic stem cell transplantation has become the most effective means to treat refractory diseases such as leukemia, severe aplastic anemia and certain malignant tumors. Since allogeneic hematopoietic stem cell transplantation requires strict tissue matching, the source of HLA-matched donors is extremely limited, which limits the wide application of hematopoietic stem cell transplantation in clinical practice. ES cells with developmental pluripotency can differentiate into hematopoietic stem cells in vitro, but ES cells have problems such as low differentiation efficiency and hematopoietic hypoplasia. It is urgent to invent ...

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Application Information

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IPC IPC(8): C12N5/0789
Inventor 那晓东李树浓赵自平
Owner SUN YAT SEN UNIV
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