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Programmed induction method for promoting embryo stem cell differentiation to hemapoietic stem cell

A technology for hematopoietic stem cells and embryonic stem cells, which is applied in the field of inducing the directional differentiation of embryonic stem cells, can solve the problems of limiting the wide application of hematopoietic stem cell transplantation, limited sources of HLA-matched donors, and low differentiation efficiency of ES cells. The effect of the patient's life and the promotion of functional development

Inactive Publication Date: 2006-02-22
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

Method used

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  • Programmed induction method for promoting embryo stem cell differentiation to hemapoietic stem cell

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Embodiment Construction

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

[0009] Isolate the yolk sac (embryo age: 8-11 days for mice / 4-6 weeks for humans) and AGM region (age of embryos: 10-12 days for mice / 4-6 weeks for humans) from mouse embryos or human aborted embryos and fetal liver (embryonic age: mouse 10-14 days / human 8-12 weeks) tissue, take adult mouse bilateral femur or extract adult bone marrow tissue, and prepare yolk sac, AGM, fetal Liver and bone marrow single cell suspension, using density gradient centrifugation method to separate mononuclear cells in DMEM containing 15% fetal bovine serum 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 identi...

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Abstract

The invention relates to a method of programmed induction for promoting directional differentiation from embryo stem cells (ES cells) to hemapoietic stem cells, which comprises the steps of tissue selection, stroma cell culturing, non-serum condition culture liquid gathering, programmed induction of differentiation from ES cells to hemapoietic stem cells, and sorting the hemapoietic stem cells.

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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/0789
Inventor 那晓东李树浓赵自平
Owner SUN YAT SEN UNIV
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