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Stem cell bottoming 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 agglomeration growth, slow growth, etc.

Inactive Publication Date: 2019-12-13
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Stem cell bottoming working solution and application thereof
  • Stem cell bottoming working solution and application thereof
  • Stem cell bottoming 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 particularly relates to a stem cell bottoming working solution and application thereof. The invention provides the stem cell bottoming working solution which comprises Matrigel and VTN-N in the volume ratio being 2:1-3:1. Experimental results indicate that the stem cell bottoming working solution comprising Matrigel and VTN-N inthe volume ratio being 2:1-3:1 is adopted for bottoming treatment and then stem cell recovery, the recovery effect is good, aggregation growth can be reduced, the growth speed can be increased, and cell death can be reduced. The stem cell bottoming working solution is adopted for bottoming treatment and recovery culture, and the number of stem cells conforms to 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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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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