Cell culture container and primary cell culture method

A cell culture and container technology, applied in the field of cell culture, can solve the problem of low yield of primary cells

Active Publication Date: 2021-01-26
BIOISLAND LAB
View PDF13 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In clinical applications, the tissue block culture method has higher safety because it does not use heterologous proteolytic enzymes to treat the cells. However, when the tissue block culture method is used for primary culture, the yield of primary cells is low

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
  • Cell culture container and primary cell culture method
  • Cell culture container and primary cell culture method
  • Cell culture container and primary cell culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The structure of the cell culture vessel used for primary culture in embodiment 1 is as follows: figure 1 shown. The culture unit of the cell culture container in embodiment 1 is one, and the material of culture tank is polystyrene, and the diameter of culture tank is 100mm, and the depth of culture tank is 20mm, and the length of fixture is 10mm, and the diameter of fixture is 1mm , the diameter of the cover plate is 92mm, the diameter of the vent hole is 0.6mm, and the diameter of the fixing hole is 1.1mm.

[0058] The steps of the primary culture of embodiment 1 are as follows:

[0059] (1) Umbilical cord collection: collect full-term healthy cesarean section fetal umbilical cord (1.5 cm in diameter), immerse in PBS containing 1% (m / v) penicillin and 1% (m / v) streptomycin, place on ice.

[0060] (2) Tissue separation: cut the umbilical cord into 4 cm long sections in a clean bench, cut them longitudinally, rinse them repeatedly with sterile PBS until the liquid is...

Embodiment 2

[0069] The primary culture steps of Example 2 are roughly the same as those of Example 1, the difference being that the tissue used in Example 2 is amnion tissue during the culture process.

[0070] According to statistics, in Example 2, the amount of cells crawling out of every 4cm of umbilical cord tissue is 6.1×10 7 The adherence rate of the tissue block was 79.9%.

[0071] Table 2 shows the results of the flow cytometric identification of the third-generation mesenchymal stem cells in Example 2.

[0072] Table 2

[0073]

[0074]

[0075] As can be seen from Table 2, the positive rate of the surface expression of CD105, CD73, CD90, CD45, CD34, CD14, CD19 and HLA-DR in the third generation mesenchymal stem cells of Example 2 is in line with international standards (specified in international standards, mesenchymal The cell surface of stromal stem cells expresses CD105, CD73 and CD90 (≥95%), and does not express CD45, CD34, CD14 or CD11b, CD79α or (≤2%)).

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

PropertyMeasurementUnit
Diameteraaaaaaaaaa
Lengthaaaaaaaaaa
Diameteraaaaaaaaaa
Login to view more

Abstract

The invention relates to a cell culture container and a primary cell culture method. The cell culture container comprises at least one culture unit, wherein the culture unit comprises a culture tank,a fixing part and a cover body, the culture tank is used for containing a cell culture solution, and the fixing part is located at the bottom of the culture tank and used for fixing a tissue block, sothat a tissue block can be attached at the bottom of the culture tank, and the cover plate can be contained in the culture tank and is movably connected with the fixing part. The cell culture container can be used for increasing the yield of primary culture cells.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cell culture vessel and a method for culturing primary cells. Background technique [0002] Primary culture refers to the first culture of tissue or cells taken out of the body, also called primary culture. Primary culture takes a short time in vitro, and its genetic properties are similar to those of cells in vivo, making it suitable for studies on cell morphology, function, and differentiation. The most commonly used primary cultures are tissue block culture and dispersed cell culture. In the tissue block culture method, the chopped tissue blocks are directly transplanted into a culture container, and then cultured after adding medium. Dispersed cell culture method is to use proteolytic enzymes (such as trypsin and collagenase) to digest the combination between cells, or use metal ion chelating agents (such as EDTA) to remove the Ca that cells rely on for mutual adhesi...

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): C12M3/04C12N5/0775
CPCC12M23/02C12M23/24C12M23/38C12M23/46C12N5/0668
Inventor 徐洪杰秦大江
Owner BIOISLAND LAB
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