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A method for preparing vascular endothelial cells and its special kit

A vascular endothelial and kit technology, applied in the field of biomedicine, can solve the problems of high price, limited amplification ability, uncertain culture medium composition, etc., and achieve the effect of high safety, stable quality and great application value

Active Publication Date: 2021-04-27
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is difficult to obtain a large number of vascular endothelial cells, and the expansion ability is limited, the price is expensive, the composition of the culture medium is uncertain, and it contains animal-derived components, which greatly limits the relevant clinical applications.

Method used

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  • A method for preparing vascular endothelial cells and its special kit
  • A method for preparing vascular endothelial cells and its special kit
  • A method for preparing vascular endothelial cells and its special kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1, the preparation of nutrient solution

[0076] 1. Preparation of stock solution

[0077] Take recombinant human serum albumin, dissolve it with sterile PBS buffer, and then filter it with a filter membrane to obtain a stock solution of recombinant human serum albumin with a concentration of 50 mg / mL.

[0078] Take human transferrin, dissolve it with sterile PBS buffer, and then filter it with a filter membrane to obtain a human transferrin stock solution with a concentration of 1 mg / mL.

[0079] Take vitamin C, dissolve it with cell culture grade water, and then filter it with a filter membrane to obtain a vitamin C stock solution with a concentration of 50 mg / mL.

[0080] Take sodium selenite, dissolve it with cell culture grade water, and then filter it with a filter membrane to obtain a sodium selenite stock solution with a concentration of 100 μg / mL.

[0081] Take CHIR-99021 and dissolve it with DMSO to obtain a stock solution of CHIR-99021 with a con...

Embodiment 2

[0093] Example 2, the acquisition and detection of vascular endothelial cells

[0094] Human pluripotent stem cells are H1 cells, H7 cells, H9 cells or CD34-iPSC cells.

[0095] The culture solution II is the culture solution II-1, the culture solution II-2 or the culture solution II-3.

[0096] The culture solution I is the culture solution I-1, the culture solution I-2, the culture solution I-3, the culture solution I-4 or the culture solution I-5.

[0097] 1. Acquisition of vascular endothelial cells

[0098] 1. Take human pluripotent stem cells grown to a confluence of 70%-80%, discard the supernatant, and then add PBS buffer to wash twice (the purpose is to remove dead cells and residual cell metabolic waste).

[0099] The morphology of human pluripotent stem cells is shown in figure 1 (The upper left is H1 cells, the upper right is H7 cells, the lower left is H9 cells, and the lower right is CD34-iPSC cells).

[0100] 2. After completing step 1, first add an appropri...

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PUM

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Abstract

The invention discloses a method for preparing vascular endothelial cells and a special kit thereof. The kit contains culture solution II and culture solution I; culture solution II includes serum albumin, transferrin, vitamin C, sodium selenite, VEGF, bFGF and serum-free basal culture solution; culture solution I includes serum Albumin, transferrin, vitamin C, sodium selenite, BMP4, GSK3 inhibitors and serum-free basal medium. Experiments have proved that vascular endothelial cells can be obtained by using the preparation method provided by the present invention, and the culture medium used in the preparation method can not only rapidly and efficiently differentiate human pluripotent stem cells into vascular endothelial cells, but also has a defined chemical composition, no animal-derived protein, No insulin added. The preparation method provided by the invention can produce human vascular endothelial cells on a large scale, with stable quality and high safety. The invention has great application value.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for preparing vascular endothelial cells and a special kit thereof. Background technique [0002] Human pluripotent stem cells (including human induced pluripotent stem cells and human embryonic stem cells) can be differentiated into functional cells of various tissues and organs, used to study human development, make disease models, and replace damaged and diseased cells through cell transplantation , Promote the body's wound repair and treat diseases. Stem cells and regenerative medicine will change the traditional treatment methods for diseases such as necrosis and injury, and bring about revolutionary changes in the mechanism research and clinical treatment of diseases. [0003] Vascular endothelial cells are important constituent cells of human blood vessels, and have important application value in injury repair, vascular reconstruction, treatment of ischemic diseases, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/069C12N2500/10C12N2500/12C12N2500/24C12N2500/38C12N2501/115C12N2501/165C12N2501/998
Inventor 那洁朱咏林张凤枝段福宇
Owner TSINGHUA UNIV
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