Method for separating living cell and constructing cell bank by means of tissue homogenate method

A technology for tissue homogenization and tissue cells, applied in chemical libraries, combinatorial chemistry, library creation, etc., can solve problems such as time-consuming, high requirements for technicians’ operating skills, and cumbersome operating procedures, so as to ensure high-quality quality and improve construction Efficiency, simple operation effect

Active Publication Date: 2013-12-25
BOYALIFE
View PDF8 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the following characteristics: 1) only need to cut the tissue into 1-1.5mm 3 The operation is relatively simple; 2) the cultivation of cell clusters takes several days to achieve the separation of cells, which takes a long time; 3) the long-term cultivation of cells has a higher possibility of contamination
[0014] It can be seen that the current so

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 for separating living cell and constructing cell bank by means of tissue homogenate method
  • Method for separating living cell and constructing cell bank by means of tissue homogenate method
  • Method for separating living cell and constructing cell bank by means of tissue homogenate method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0047] Example 1: Using tissue homogenization method to separate placental lobular cells to construct a cell bank

[0048] The method of placental lobular tissue homogenization includes the following steps:

[0049] (1) Placental tissue cleaning: The placental tissue is processed in a biological safety cabinet. According to the size of the placenta, use an appropriate amount of PBS buffer containing 1% bi-antibody to wash the placental tissue until the residual blood on the surface of the placental tissue is washed away, and the placental surface is free. Blood clot;

[0050] (2) Preliminary treatment of placental lobule tissue: Use surgical scissors to cut the placental lobules from the placental tissue obtained in step (1), transfer the placental lobules to a petri dish and cut into small pieces, and take 60-90 grams of placental lobule tissue Put the block into the homogenization container of the tissue cell processor PlacentaPro, add 50-80 ml of PBS and seal the processing cup; ...

Example Embodiment

[0060] Example 2: Comparative experiment of placental lobular tissue homogenization treatment and manual treatment

[0061] The comparison experiment was divided into 8 groups. Refer to the method of Example 1 for a fully automated tissue processing experiment. Refer to the method described in patent CN201210292509.9 (publication number CN102807966A) for manual processing (digestion) experiments. The placenta tissue is sourced from 8 different donors, and the processing volume is between 60-90 grams. The comparative experimental results are shown in Table 2.

[0062]

[0063] Table 2 Comparison of placental lobular tissue data between fully automated and manual processing

[0064] The results showed that the average cell mass per gram of placental lobules after being processed by the automatic tissue cell processor PlacentaPro was 2.4 times that of manual processing, which was significantly higher than the cells obtained by manual digestion, providing a large number of cells ...

Example Embodiment

[0065] Example 3: Primary growth of cells isolated from placental lobule tissue by homogenization method

[0066] (1) Preparation before cell culture: Add red blood cell lysate (Roche) at a ratio of 1 part cell suspension to 2-3 parts red blood cell lysate, incubate at 15-25°C for 10-15 minutes, and put it in the centrifuge Centrifuge at 1400 rpm for 10 minutes, remove the supernatant, observe the lysis of red blood cells, repeat this step for lysis if necessary. After lysis, add PBS to resuspend the cells and extract a small amount of samples for cell count, put them in a centrifuge at 1400 rpm for 10 minutes to wash the cells, remove the supernatant, and add mesenchymal stem cell culture medium (15% FBS+1) %L-Glutamine+0.05%Gentamicin+84%DMEM-F12) resuspend the cells, and 2-5×10 4 / Cm 2 Placental-derived nucleated cells are inoculated into the culture flask at a density of 5%.

[0067] (2) Primary cell culture: put the culture flask into C0 2 The culture is carried out in an inc...

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 for separating the tissue cell of a fetal appendage and constructing a cell bank by means of a tissue homogenate method, so that the process for constructing the cell blank is simple in steps, time-saving, and capable of achieving easy quality control and standardization. The method disclosed by the invention comprises the following steps of (1) cleaning the tissue of the fetal appendage; (2) carrying out homogenate pretreatment on the tissue; (3) carrying out homogenate treatment on the tissue to separate a cell (cell cluster); (4) filtering the tissue and collecting the cell (cell cluster) after the homogenate; (5) detecting the cell (cell cluster); (6) cryopreserving the separated cell (cell cluster) and constructing the cell bank. Compared with the method in the prior art, the method disclosed by the invention has the advantages of simple operation, short time consumption, no introduction of an exogenous reagent, easiness for standardization, easy quality control, great cell gain and the like, and is of great significance in improving the construction efficiency of the fetal appendage cell bank and obtaining the stem cell which is high in quality and applied to treatment.

Description

technical field [0001] The invention relates to a method for constructing a fetal appendage tissue cell bank, more specifically a physical method for separating cells from the fetal appendage tissue to construct a cell (cluster) bank. Background technique [0002] Fetal appendage tissue is mainly composed of placental lobules, placental base, decidua basalis, amniotic membrane and umbilical cord tissue. Originating from the extraembryonic mesoderm during embryonic development, the placenta is composed of mesenchyme, blood vessels and trophoblast cells. It is the earliest hematopoietic organ of the fetus and contains a large amount of mesenchyme. The latest research shows that the placenta contains mesenchymal stem cells, placental sub-totipotent stem cells and abundant hematopoietic stem cells. The isolation and culture of these pluripotent stem cells from the placenta will open up a new and rich source for experimental research and clinical application. [0003] Placental ...

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): C40B50/06
Inventor 许晓椿肖海蓉
Owner BOYALIFE
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