Method of harvesting cells on three-dimensional microcarrier

A microcarrier, three-dimensional technology, applied in the field of harvesting cells on three-dimensional microcarriers, can solve problems such as cell damage, and achieve the effect of mild cells and maintaining cell activity.

Active Publication Date: 2019-04-16
BEIJING CYTONICHE BIOTECH CO LTD
View PDF2 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, after three-dimensional cell culture, multi-layered or agglomerated cells are difficult to be digested by trypsin, and the long-term digestion of trypsin or its substitutes will cause great damage to the cells themselves. Therefore, a more gentle method is needed. Methods Harvesting cells from the three-dimensional carrier, that is, by cracking the three-dimensional carrier material without damaging the cells, the efficient harvesting of cells from the three-dimensional carrier material can be achieved, thereby meeting the needs of subsequent applications
For cell culture on 3D microcarriers, there is currently no method for more gentle harvesting of cells on this carrier for subsequent applications

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method of harvesting cells on three-dimensional microcarrier
  • Method of harvesting cells on three-dimensional microcarrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Lysis solution formula: 0.1% collagenase, 0.1% ethylenediaminetetraacetic acid, 0.05% trypsin; stop solution: full medium

[0030] 2. Transfer 20mg of three-dimensional microcarriers with adipose-derived mesenchymal stem cells from the culture dish to a centrifuge tube, centrifuge at 400×g for 2 minutes, drain the supernatant, add an appropriate amount of PBS, and shake gently by hand for 20-30s , aspirate the supernatant as much as possible, and repeat the PBS wash once;

[0031] 3. Add 3mL of lysate and incubate in a cell culture incubator at 37°C for 30min. During this period, gently pipette several times with a 1ml pipette every 10min;

[0032] 4. After 30 minutes, the microcarriers are completely lysed, and 3 mL of full medium is added to terminate the lysis process;

[0033] 5. Centrifuge at 200×g for 5 minutes, discard the supernatant, and resuspend the cells according to the requirements of subsequent applications.

[0034] 6. Cell detection method:

[003...

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 discloses a method of harvesting cells on a three-dimensional microcarrier. The method comprises the following steps of 1, executing natural sedimentation or centrifugation of a three-dimensional microcarrier adsorbing cultured cells, and removing the supernatant to obtain a three-dimensional microcarrier containing cells; 2, adding the three-dimensional microcarrier containing the cells into lysate, and incubating and lysing the three-dimensional microcarrier; 3, after the three-dimensional microcarrier in step 2 is completely lysed, a stop buffer is added to terminate the lysisprocess or directly executing the step 4 to terminate the lysis; 4, centrifuging the system obtained in step 3, and discarding the supernatant to obtain the cells on the three-dimensional microcarrier. The cells on the carrier can be more gently obtained for subsequent application; by lysing the microcarrier, the microcarrier is dissolved from a solid state to a liquid state, accordingly the cells on the carrier are released to the solution, and the solution and the cells are separated in a centrifugation way so as to obtain the cells.

Description

technical field [0001] The invention relates to a method for harvesting cells on three-dimensional microcarriers, belonging to the technical field of cell culture. Background technique [0002] With the rapid development of interdisciplinary subjects such as biomedicine, materials science, mechanics, and engineering, more and more people are paying attention to the difference between the microenvironment of cells cultured in vitro and the microenvironment in vivo. The traditional two-dimensional cell culture (based on commercial culture dishes or multi-well plates) technology has been developed for more than 100 years, and it has been widely used in basic life science research, medical research and other fields. However, with the rapid development of microscopic imaging technology, researchers have gradually discovered the living state of cells in a two-dimensional culture environment, and the cell morphology is far from the culture conditions in the real environment in vivo...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/0075
Inventor 鄢晓君刘伟
Owner BEIJING CYTONICHE BIOTECH CO LTD
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