A kind of stem cell base layer working solution and application thereof

A stem cell and working solution technology, applied in the field of stem cell culture, can solve the problems of high mortality, easy aggregation growth, slow growth, etc., and achieve the effect of reducing cell death, accelerating growth speed, and good recovery effect

Inactive Publication Date: 2021-05-11
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of this, the present invention provides a stem cell primer working solution and its application, which are used to solve the problems in the prior art that the resuscitated or cultured stem cells are prone to aggregate growth, slow growth or high mortality

Method used

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  • A kind of stem cell base layer working solution and application thereof
  • A kind of stem cell base layer working solution and application thereof
  • A kind of stem cell base layer working solution and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1) Prepare different groups of stem cell primer working solutions

[0040] Freeze and thaw Matrigel and VTN-N from -80°C to 4°C overnight, and add 100 μL Matrigel and VTN-N to 10ml DPBS in a 15mL centrifuge tube to dilute at a ratio of 1:100 the next day to obtain Matrigel dilution and VTN-N dilution, stored in 4 ℃ refrigerator for future use.

[0041] Use Matrigel diluent and VTN-N diluent to prepare different groups of stem cell priming working solution. The total volume of stem cell priming working solution is 2mL, as follows:

[0042] Group 1: Matrigel diluent 2mL;

[0043] Group 2: VTN-N dilution 2mL;

[0044] Group 3: Matrigel diluent 1mL + VTN-N diluent 1mL;

[0045] Group 4: Matrigel diluent 1.33mL+VTN-N diluent 0.67mL;

[0046] Group 5: Matrigel diluent 1.5mL + VTN-N diluent 0.5mL;

[0047] Group 6: Matrigel diluent 0.67mL + VTN-N diluent 1.33mL;

[0048] Group 7: 0.5 mL of Matrigel diluent + 1.5 mL of VTN-N diluent.

[0049] 2) Bottom treatment of the p...

Embodiment 2

[0061] For the three groups of hiPSCs, use the working solution of group 5 stem cells for bottoming, and then recover and culture them in the same way as in Example 1, and count the number of hiPSCs for three days. The results are as follows Figure 8 as shown, Figure 8 In Example 2 of the present invention, hiPSCs were primed with the stem cell working solution of group 5 and then resuscitated. The results showed that the growth of hiPSCs was consistent with the growth of stem cells Trend, the stem cell base working solution of the present invention has effectiveness and feasibility.

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Abstract

The invention belongs to the technical field of stem cell culture, and in particular relates to a stem cell priming working solution and an application thereof. The invention provides a working solution for stem cell priming, comprising: Matrigel and VTN-N; the volume ratio of the Matrigel to the VTN-N is 2:1-3:1. The experimental results show that using the stem cell priming working solution with a volume ratio of Matrigel and VTN‑N of 2:1 to 3:1 for priming and then resuscitating stem cells has a good recovery effect, can reduce aggregate growth, accelerate growth speed, and reduce cell death ; Using the stem cell bottoming working solution of the present invention to carry out the bottoming treatment and then resuscitating culture, the number of stem cells grows in line with the growth trend of stem cells.

Description

technical field [0001] The invention belongs to the technical field of stem cell culture, and in particular relates to a stem cell priming working solution and an application thereof. Background technique [0002] Embryonic stem cells (ESCs) can differentiate into all cells derived from the three germ layers in vivo, and then participate in the formation of all tissues and organs in the body. They are of great application value in the fields of regenerative medicine, tissue engineering, and drug discovery and evaluation. However, the ethics, immune rejection and insufficient resources faced by ESCs hinder its development in the basic research and clinical application of stem cell biology, while the acquisition method of human induced pluripotent stem cells (hiPSC) can be very good. overcame these problems. Therefore, in the field also called "stem cells", compared with embryonic stem cells, hiPSCs can not only avoid physiological and ethical problems, but also avoid uncontr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074
CPCC12N5/0696C12N2533/50C12N2533/90
Inventor 苗小敏汤亚东梁大锡罗竣仁周颖陈巧桐李卓刚陈家盈张焜
Owner GUANGDONG UNIV OF TECH
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