Differential culture medium of human pluripotent stem cell derived mesoderm cells and application of differential culture medium

A technology of human pluripotent stem cells and differentiation medium, which is applied in the field of cell differentiation culture, can solve the problems of low differentiation efficiency, low grade of raw materials, and high cost, and achieve the effects of high differentiation efficiency, clear components, and high universality

Active Publication Date: 2021-03-26
BEIJING TRANSGEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the human pluripotent stem cell-derived mesoderm cell differentiation medium has the following deficiencies: (1) The raw material grade is low
The grades of raw materials are all biological and animal-derived ingredients, and it is difficult to meet the quality control requirements for raw materials of cell therapy products
(2) High cost
Using STEMdiff TM When Mesoderm Induction Medium (product number #05220) is used to prepare mesoderm cells derived from human pluripotent stem cells, only the differentiation medium is counted as a raw material, and the production cost of raw drug cells has reached as high as 800 yuan / 1.0×10 7 Brachyury(T) + OCT4 - Mesoderm cells derived from human pluripotent stem cells are difficult to achieve inclusive cell therapy products
(3) Poor stability between batches
(4) Low differentiation efficiency
These problems largely limit the industrial transformation and clinical application of functional cells derived from human pluripotent stem cells

Method used

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  • Differential culture medium of human pluripotent stem cell derived mesoderm cells and application of differential culture medium
  • Differential culture medium of human pluripotent stem cell derived mesoderm cells and application of differential culture medium
  • Differential culture medium of human pluripotent stem cell derived mesoderm cells and application of differential culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1 Differentiation medium for mesoderm cells derived from human pluripotent stem cells

[0088] A differentiation medium for human pluripotent stem cell-derived mesoderm cells, the differentiation medium consists of the following components:

[0089] Basal medium: RPMI1640 medium (1×);

[0090] Compound combination: the components and final use concentrations of the compound combination are: D-pipercolic acid 300 μg / L, triiodo-L-thyronine 800 μg / L and thiamine hydrochloride 45 mg / L;

[0091] GSK-3 inhibitor: the GSK-3 inhibitor and the final use concentration are CHIR99021 8 μM.

Embodiment 2

[0092] Example 2 Differentiation medium for mesoderm cells derived from human pluripotent stem cells

[0093] A differentiation medium for human pluripotent stem cell-derived mesoderm cells, the differentiation medium consists of the following components:

[0094] Basal medium: DMEM / F12 medium (1×);

[0095] Compound combination: the components and final use concentrations of the compound combination are: D-pipercolic acid 300 μg / L, triiodo-L-thyronine 800 μg / L and thiamine hydrochloride 45 mg / L;

[0096] GSK-3 inhibitor: the GSK-3 inhibitor and the final use concentration are CHIR99021 8 μM.

Embodiment 3

[0097] Example 3 Differentiation medium for human pluripotent stem cell-derived mesoderm cells

[0098] A differentiation medium for human pluripotent stem cell-derived mesoderm cells, the differentiation medium consists of the following components:

[0099] Basal medium: IMDM medium (1×);

[0100] Compound combination: the components and final use concentrations of the compound combination are: D-pipercolic acid 300 μg / L, triiodo-L-thyronine 800 μg / L and thiamine hydrochloride 45 mg / L;

[0101] GSK-3 inhibitor: the GSK-3 inhibitor and the final use concentration are CHIR99021 8 μM.

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Abstract

The invention discloses a differentiation culture medium of human pluripotent stem cell derived mesoderm cells and application of the differentiation culture medium. The invention firstly discloses the differentiation culture medium of human pluripotent stem cell derived mesoderm cells. The differentiation culture medium comprises the following components of a basal culture medium, a compound combination and a glycogen synthase kinase 3 (GSK-3) inhibitor, wherein the compound combination comprises the following components of D-piperidine acid, triiodo-L-thyronine and thiamine hydrochloride. The invention further discloses application of the differentiation culture medium in inducing differentiation of the human pluripotent stem cells into mesoderm cells and / or mesoderm downstream derived cells. The differentiation culture medium for the pluripotent stem cell derived mesoderm cells is high in raw material level and low in cost, is high in batch-to-batch stability, high in differentiation efficiency and high in universality when used for inducing the human pluripotent stem cells to differentiate into the mesoderm cells, and has very high industrial transformation and clinical application values.

Description

technical field [0001] The invention relates to the technical field of cell differentiation and culture. More specifically, it relates to a differentiation medium for human pluripotent stem cell-derived mesoderm cells and applications thereof. Background technique [0002] Human pluripotent stem cells (hPSCs) include embryonic stem cells (humanembyonic stem cells, hESCs) and induced pluripotent stem cells (human induced pluripotent stem cells, hiPSCs), which can proliferate infinitely in vitro while maintaining "stemness". And can differentiate into almost all types of somatic cells of the body. Human pluripotent stem cells can further differentiate into functional cells such as cardiomyocytes, mesenchymal stem cells, hematopoietic stem cells, and T cells through mesoderm cells. These functional cells can supplement or replace different diseased tissue cells, and are used to treat diseases and promote tissue repair and regeneration, and have important regenerative medical ...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775C12N5/0789C12N5/0735C12N5/10
CPCC12N5/0647C12N5/0657C12N5/0662C12N2500/38C12N2501/395C12N2501/727C12N2501/999C12N2506/02C12N2506/45C12N2510/00
Inventor 辛文佟曼韩伟方陆曼薛博涵马静
Owner BEIJING TRANSGEN BIOTECH CO LTD
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