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Method for preparing stem cell active factor

A technology of active factors and stem cells, applied in the field of stem cells, can solve the problems of cytokine loss, secondary pollution of stem cells, short storage time, etc.

Active Publication Date: 2020-06-23
BOYALIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the liquid culture supernatant has a short storage time at room temperature, which hinders its promotion and application.
[0008] In addition, for the current stem cell sorting technology, magnetic bead sorting is more common. However, in conventional sorting, the magnetic beads will be cultured together with the stem cells, that is, the magnetic beads are not detached from the stem cells. The later production will definitely have a certain impact. Although there are some methods, such as adding some bacteria or other substances to degrade, this is equivalent to adding other exogenous proteins and other components, causing secondary pollution to stem cells, so doesn't actually solve the above problem
[0009] There are also some reports on the extraction and purification of stem cell culture supernatant, but the effect is still not good. For example, the patent with the notification number CN102600057B discloses a preparation method of human placental stem cell extract freeze-dried powder. A single 3000D filter membrane is used for ultrafiltration interception, which removes some impurities, but the impurities such as sugar are still not removed, and it also causes a large loss of cytokines, poor repair effect, and also causes discomfort such as dry skin , the practical application value is low
[0010] In addition, it is currently difficult for people to prepare stem cell active factors with high efficiency and / or stable performance

Method used

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  • Method for preparing stem cell active factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1: Preparation of Stem Cell Active Factor Concentrate

[0088] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:

[0089]The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) Serum-free medium, cultured in a 37°C, 5% CO2 incubator, cell colonies appear after 7 days of culture; cells reach saturation in about 12 days, and can be subcultured; the cells are long spindle-shaped, which is a typical fibroblast Cell-like morphology (see for example figure 1 );

[0090] (2) Cell expansion:

[0091] When the cell confluency reaches 70-80%, gently wash the cells in the culture flask with physiological saline 1-2 times, add 4ml of digestion solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe...

Embodiment 2

[0100] Embodiment 2: Preparation of stem cell active factor concentrate

[0101] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:

[0102] The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) Serum-free medium, cultured in 37°C, 5% CO2 incubator, cell colonies appear after 7 days of culture; cells reach saturation in about 10 days, and can be subcultured; cells are long spindle-shaped, which is a typical fibroblast cell-like morphology;

[0103] (2) Cell expansion:

[0104] When the cell confluency reaches 70-80%, gently wash the cells in the culture flask with physiological saline for 1-2 times, add 5ml of digestive solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe under the microscope that ...

Embodiment 3

[0112] Embodiment 3: Preparation of stem cell active factor concentrate

[0113] (1) Acquisition of umbilical cord-derived mesenchymal stem cells:

[0114] The umbilical cord of a healthy full-term cesarean section was taken, and prepared by separating Wharton's Jelly (Wharton's Jelly) and using the Wharton's Jelly suspension method; put Wharton's Jelly in a T75 culture bottle, and add an appropriate amount of mesenchymal stem cells (MSC ) serum-free medium, cultured in a 37°C, 5% CO2 incubator, and cell colonies appeared after 7 days of culture; the cells reached saturation in about 13 days and could be subcultured; the cells were long spindle-shaped, which is a typical fibroblast cell-like morphology;

[0115] (2) Cell expansion:

[0116] When the cell confluence reaches 70-80%, gently wash the cells in the culture flask with physiological saline for 1-2 times, add 3ml of digestion solution (0.25% trypsin), and leave it at room temperature for 1-2 minutes. Observe under ...

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Abstract

The invention relates to a method for preparing a stem cell active factor, and on one hand, in particular relates to a method for preparing the stem cell active factor. The method comprises the following steps of: (1) acquiring umbilical cord placenta derived mesenchymal stem cells; (2) performing cell amplification; (3) performing cell identification detection; (4) preparing the stem cell active factor; and (5) performing optional freeze-drying. Furthermore, the invention relates to a stem cell active factor freeze-dried powder agent prepared by using the method demonstratively. The method provided by the invention has the beneficial effects as shown in the specification of the invention.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a human-derived mesenchymal stem cell factor and its preparation method and application, in particular to a method for preparing stem cell active factors from umbilical cord mesenchymal stem cells. The method of the invention can prepare stem cell active factors with high efficiency and stable performance. Background technique [0002] In recent years, with the deepening of research on mesenchymal stem cells, its secreted factors have become a hot spot for researchers. Mesenchymal stem cells can secrete a variety of biologically active cytokines, which can effectively regulate body cell signal transduction, activate human stem cells, and then physiologically repair or replace damaged, diseased and aging cells in the body. [0003] The application of stem cells in cosmetology is the earliest use of stem cells in plastic surgery. It is a stem cell cosmetology techno...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775C12P21/00C07K1/34A61K8/64A61K8/98A61Q19/00A61Q19/02
CPCA61K8/64A61K8/981A61Q19/00A61Q19/02C07K1/34C12N5/0668C12P21/00
Inventor 王芳周丹肖海蓉
Owner BOYALIFE
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