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Method for inducing and differentiating human multipotent stem cells into aged blood cells

A technology for inducing human pluripotent stem cells and inducing differentiation is applied in the field of establishing human pluripotent stem cells to induce differentiation into mature blood cells. desired effect

Inactive Publication Date: 2017-06-13
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Solve the problems of cumbersome induction technology, high cost, low output and potential safety hazards in the existing technology

Method used

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  • Method for inducing and differentiating human multipotent stem cells into aged blood cells
  • Method for inducing and differentiating human multipotent stem cells into aged blood cells
  • Method for inducing and differentiating human multipotent stem cells into aged blood cells

Examples

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Effect test

Embodiment 1

[0054] A method for establishing human pluripotent stem cells induced to differentiate into mature blood cells, including two steps (the flow chart of the method is shown in figure 1 Shown): A. Human pluripotent stem cells are induced to differentiate into CD31 and CD34 hematopoietic progenitor cells; B. CD31 and CD34 hematopoietic progenitor cells are induced to differentiate into human mature blood cells; the specific operations are as follows:

[0055] A. Human pluripotent stem cells were induced to differentiate into hematopoietic progenitor cells of CD31 and CD34

[0056] (1) Culture iPS cells in a 6-well plate using Matrigel-based mTeSR1 medium without a feeder layer (Stemcell Technologies), replace the mTeSR1 medium every day, and culture for 4 days, and the cell volume per well reaches 1.5-2.5 ×10 6 , and then digest the cells. When digesting the cells, discard the mTeSR1 medium, wash it with 1mL cell digestion buffer (Cell Dissociation Buffer, referred to as CDB, Gibc...

Embodiment 2

[0077] A method for establishing human pluripotent stem cells induced to differentiate into mature blood cells, the specific operations are as follows:

[0078] A. Human pluripotent stem cells were induced to differentiate into hematopoietic progenitor cells of CD31 and CD34

[0079] (1) Culture hES cells in a 6-well plate using the feeder-free Matrigel-based mTeSR1 medium, replace the mTeSR1 medium every day, culture for 5 days, and the cell volume per well reaches 1.5-2.5 × 10 6 , and then digest the cells. When digesting the cells, discard the mTeSR1 medium, wash it once with 1mL cell dissociation buffer (CDB, Gibco company for short), discard the cell dissociation buffer, then add 1mL cell dissociation buffer, Incubate in a 37°C incubator for 8 minutes. When the cell clones are separated and loose, remove the cell digestion solution, add 1-2mL mTeSR1 medium to each well, and the cells appear to be in a floating state;

[0080] (2) Transfer the cells to a centrifuge tube a...

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Abstract

The invention belongs to the field of induced differentiation of blood cells, and provides a method for inducing and differentiating human multipotent stem cells into aged blood cells. The method comprises the following steps of A, inducing and differentiating the human multipotent stem cells into hematopoietic progenitor cells CD31 and CD34; B, inducing and differentiating the hematopoietic progenitor cells CD31 and CD34 into human aged cells. The method has the advantages that by adopting the two-step inducing type of single-layer cell culture and suspension cell culture, the inducing procedure is simplified, the inducting time is shortened, the operation is simple, the efficiency is high, and the stability is realized; by adopting the inducing and differentiating system without feed layer cells, the induced blood cell has biological safety guarantee, the requirements of clinical conversion and clinical experiment can be met, and the medical conversion value is high.

Description

technical field [0001] The invention belongs to the field of induction and differentiation of blood cells, and in particular relates to a method for establishing human pluripotent stem cells to induce differentiation into mature blood cells. Background technique [0002] Blood transfusion is the main treatment for severe congenital and acquired anemia diseases, including hematological malignancies, red blood cell aplasia anemia, thalassemia, hemoglobinopathies, etc., which seriously threaten human life and health. However, insufficient matching and scarcity of blood substitutes still limit the treatment of these diseases. In recent years, with the technical breakthrough of hematopoietic progenitor / stem cells and CD34-positive cells successfully differentiated into red blood cells, it has brought new hope for the treatment of blood diseases, but there are still problems such as difficulty in matching. [0003] Human blood cells, especially red blood cells, cannot expand in v...

Claims

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Application Information

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IPC IPC(8): C12N5/078C12N5/10C12N5/0735
CPCC12N5/0634C12N2501/11C12N2501/115C12N2501/155C12N2501/727C12N2506/02C12N2506/45
Inventor 王华岩卢世江王宁
Owner NORTHWEST A & F UNIV
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