Preparation method of high-concentration human-source mesenchymal stem cell lysis solution

A high-concentration stem cell technology, applied in the field of preparation of high-concentration human mesenchymal stem cell lysate, can solve the problems of low purity of stem cell bioactive factors, difficulty in removing tissue proteins, and low concentration of active factors, and achieve proliferation and differentiation Strong ability, stable biological properties, and the effect of repairing damaged cells

Active Publication Date: 2020-02-18
深圳市润科生物科技有限公司
View PDF24 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the method for extracting stem cell bioactive factors in practice has the following deficiencies: (1) direct use of tissues such as placenta to extract, the obtained active protein contains a large amount of tissue protein that is difficult to remove, and the obtained stem cell bioactive factors have low purity; (2) Utilizing stem cell expansion culture fluid to extract active factors requires concentrating a large amount of culture fluid, whi

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for preparing a high-concentration human-derived mesenchymal stem cell lysate, comprising the following steps:

[0041] 1. Breaking step: Use 60W ultrasonic waves to ultrasonically break the collected human umbilical cord mesenchymal stem cells with a fusion degree of 80%. The ultrasonic waves are applied alternately for 1 second and stopped for 2 seconds. The total breaking time is 30 minutes to obtain broken cell fluid.

[0042] 2. Centrifugation step: the broken cell solution was centrifuged for 30 min under the conditions of a temperature of 2° C. and a centrifugal force of 2000 g, and the supernatant was obtained after centrifugation.

[0043] 3. Purification step: use a 3K ultrafiltration membrane to filter and concentrate the supernatant to obtain a concentrated solution whose volume is 1 / 5 of the volume of the supernatant; use a hollow fiber ultrafiltration column with a molecular weight cut-off of 100KD to purify the supernatant. The concentrated solutio...

Embodiment 2

[0045] A method for preparing a high-concentration human-derived mesenchymal stem cell lysate, comprising the following steps:

[0046] 1. Breaking step: Use 100W ultrasonic waves to ultrasonically break the collected human umbilical cord mesenchymal stem cells with a fusion degree of 90%. The ultrasonic waves are applied alternately for 5 seconds and stopped for 6 seconds. The total breaking time is 10 minutes to obtain broken cell liquid.

[0047] 2. Centrifugation step: centrifuge the broken cell liquid for 10 min under the condition of temperature of 8° C. and centrifugal force of 5000 g, and take the supernatant after centrifugation.

[0048] 3. Purification step: use a 3K ultrafiltration membrane to filter and concentrate the supernatant to obtain a concentrated solution with a volume of 1 / 8 of the volume of the supernatant; use a hollow fiber ultrafiltration column with a molecular weight cut-off of 100KD to filter the supernatant. The concentrated solution is further c...

Embodiment 3

[0050] A method for preparing a high-concentration human-derived mesenchymal stem cell lysate, comprising the following steps:

[0051] 1. Breaking step: Use 80W ultrasonic waves to ultrasonically break the collected human umbilical cord mesenchymal stem cells with a fusion degree of 85%. The ultrasonic waves are applied alternately for 3 seconds and stopped for 4 seconds. The total breaking time is 20 minutes to obtain broken cell liquid.

[0052] 2. Centrifugation step: the broken cell solution was centrifuged for 10 min under the conditions of a temperature of 4° C. and a centrifugal force of 4000 g, and the supernatant was obtained after centrifugation.

[0053] 3. Purification step: use a 3K ultrafiltration membrane to filter and concentrate the supernatant to obtain a concentrated solution whose volume is 1 / 10 of the volume of the supernatant; use a hollow fiber ultrafiltration column with a molecular weight cut-off of 100KD to purify the supernatant. The concentrated so...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a preparation method of a high-concentration human-source mesenchymal stem cell lysis solution. The preparation method comprises the following steps of step of breaking: performing ultrasonic breaking on a human-source mesenchymal stem cell lysis solution of which the fusion degree is 80-90%, which is obtained by collection by using 60-100W ultrasonic waves for 10-30min to obtain broken cell liquid; step of centrifugation: performing centrifugation on the broken cell liquid under the condition that the temperature is 2-8 DEG C and centrifugal force is 2000-5000g for 10-30min, and after centrifugation, taking supernatant; step of purification: performing filtering and concentrating on the supernatant by a 3K ultra filtration membrane to obtain a concentrated solutionof which the volume is 1/5-1/10 of the volume of the supernatant; and performing further concentrating on the concentrated solution by a hollow fiber ultrafiltration column to obtain the high-concentration human-source mesenchymal stem cell lysis solution. The method solves the problems that in the prior art, the extracted stem cell factors are low in purity, low in yield, low in activity, low inefficiency, low in yield and the like, and the obtained stem cell lysis solution has favorable capacity for promoting proliferation of mesenchymal stem cells and promoting collagen synthesis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing a high-concentration human-derived mesenchymal stem cell lysate. Background technique [0002] Stem cells, as emerging seed cells, are cells with multi-directional differentiation potential. Transplanted stem cells can self-renew and differentiate into various cells to repair tissues; and stem cells can produce or secrete a large number of biologically active factors. These biologically active Functionally, the factors can be divided into six categories: immune regulation, anti-apoptosis, angiogenesis, support for proliferation and differentiation of stem / progenitor cells, chemotaxis, and anti-scarring. Through autocrine or paracrine methods, they improve the local microenvironment, promote the migration and differentiation of endogenous stem cells to the wound site, and then physiologically repair or replace the damaged, diseased and aging cells of the body. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N13/00C12N5/0775
CPCC12N5/0665C12N13/00C12N2509/00
Inventor 马冬磊林科佳魏宗科张若楠
Owner 深圳市润科生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap