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Cell culture solution and application thereof

A cell culture and stem cell technology, applied in the field of cell culture fluid, can solve the problems of many uncontrollable factors, high cost, high risk, etc., and achieve the effect of promoting proliferation efficiency and normal morphology

Inactive Publication Date: 2019-12-10
山东佰鸿干细胞生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The problem with the above-mentioned serum-free medium is that after a large number of human umbilical cord mesenchymal stem cells are subcultured, some human umbilical cord mesenchymal stem cells can still maintain their pluripotency, while some cells begin to age, such as the growth rate gradually slows down, The cell shape is also changed from long spindles to flat and thick, thus losing its clinical value
Moreover, the composition of the above-mentioned serum-free medium is complex, and there are many uncontrollable factors. If it is applied on a large scale, the risk will be greater and the cost will be higher.

Method used

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  • Cell culture solution and application thereof
  • Cell culture solution and application thereof
  • Cell culture solution and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0055] This embodiment provides a cell culture solution, the cell culture solution includes apoptotic bodies produced by inducing apoptosis of stem cells in vitro, and the stem cells are umbilical cord mesenchymal stem cells. In some embodiments, the stem cells are bone marrow mesenchymal stem cells, adipose mesenchymal stem cells, dental pulp stem cells, periodontal ligament stem cells, dental follicle stem cells, tooth germ progenitor cells, stem cells from apical papilla, oral epithelium Progenitor / stem cells, gingival-derived mesenchymal stem cells, periosteum-derived stem cells, apoptotic salivary gland-derived stem cells, or any combination thereof.

[0056] The amount of apoptotic bodies added to the cell culture solution described in this example was 2 μg / L. In some embodiments, the amount of apoptotic bodies added in the cell culture medium is 5 μg / L, 10 μg / L, 20 μg / L, 30 μg / L, 40 μg / L, 50 μg / L, 60 μg / L, 70 μg / L L, 80μg / L, 90μg / L, 100μg / L, 110μg / L, 120μg / L, 130μg / L, ...

Embodiment 2

[0077] This example provides the application of the cell culture solution described in Example 1 in maintaining large-scale cultivation of stem cells. The specific application process is as follows:

[0078] Cell inoculation: take well-grown mesenchymal stem cells of the 3rd to 20th generation, centrifuge, collect the cell pellet, add the cell culture solution described in Example 1 of the present invention to make a cell suspension, and count them as experimental group 1; Well-grown mesenchymal stem cells of the 20th generation were centrifuged, and the cell pellet was collected, and the culture medium disclosed in patent CN201811341164.5 was added to make a cell suspension, and then counted, which was used as control group 1. According to the results of cell counting, respectively, according to 5 × 10 3 cells / mL for subculture and inoculated in 24-well plates, 1 ml / well. Put in 5% CO after inoculation 2 , 37°C incubator for cultivation. Cell counting: 24 hours after cell ...

Embodiment 3

[0080] Example 3 Verification of osteogenic differentiation function of stem cells obtained after large-scale culture using the cell culture medium provided in the examples of the present invention

[0081]The umbilical cord mesenchymal stem cells cultured in Experimental Group 1 in Example 2 were taken, digested, centrifuged to collect the precipitate, washed with normal saline, and cell suspension was prepared as Experimental Group 2. The umbilical cord mesenchymal stem cells cultured in the control group 1 of Example 2 were taken, digested, centrifuged to collect the precipitate, washed with physiological saline, and prepared into a cell suspension, which was used as the control group 2. After counting, the cells were seeded in a six-well plate, about 2.5×10 per well 5 cells, when the cells grow to 75% ~ 85%, the liquid is replaced with an osteogenic induction solution to continue culturing, wherein the components of the osteogenesis induction solution include 500 parts by ...

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Abstract

The invention provides a cell culture solution. The cell culture solution comprises an apoptotic body, can be used for promoting the reproduction efficiency of stem cells, especially mesenchymal stemcells, and large-scale cultivation of the mesenchymal stem cells and enables the mesenchymal stem cells to have normal form and uniformity after large-scale cultivation, and can be used for keeping multipotency of the mesenchymal stem cells after large-scale cultivation, so that large-scale clinical application of the mesenchymal stem cells becomes possible.

Description

technical field [0001] The invention belongs to the technical field of bioengineering. In particular, it relates to a cell culture medium suitable for large-scale cell culture and its application. Background technique [0002] Mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) are a type of pluripotent stem cells with self-renewal and multiple differentiation potentials. After induction, they can differentiate into various tissues and cells. It is an ideal seed cell in regenerative medicine. It is worth noting that after a large number of subcultures or high-passage cultures, some cells can still maintain their pluripotency, while some cells begin to age or their differentiation function begins to weaken, that is, the growth rate gradually slows down, and the cell shape changes from long to small. The spindle becomes flat and thick, and then loses its clinical use value, and the mesenchymal stem cells used for cell therapy not only deteriorate in function but also signi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/80
Inventor 蒲锋星程蕊苹高迎凤
Owner 山东佰鸿干细胞生物技术有限公司
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