Rat II type primary alveolar epithelial cell extraction method

A technology of alveolar epithelium and extraction method, which is applied in cell dissociation methods, respiratory/lung cells, animal cells, etc., can solve the problems of waste of digestive juice, low utilization rate of digestive juice, and long time consumption, so as to increase digestion efficiency and utilization The effect of high efficiency, stable and rapid digestion, and shortened total digestion time

Inactive Publication Date: 2018-11-16
AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the applicant found in actual operation that the water bath digestion process in the traditional culture method takes a long time, and after the first injection of digestive juice, the digestive juice will continue to leak out of the lungs, so it cannot be guaranteed that there is always digestive juice in all alveoli, resulting in alveolar The epithelial cells in the alveoli are not fully digested, which reduces the digestion effect and efficiency; in order to fully digest the epithelial cells in the alveoli, the traditional method will add digestive juice to the lungs every 5 minutes, and this method will invisibly increase The amount of digestive juice is not enough, and the supplement of digestive juice is fast replenishment. During the period from the replenishment to the next replenishment, the digestive juice will continue to leak out of the lungs, and the epithelial cells in the alveoli are still not fully digested continuously; in addition, Due to the continuous replenishment of digestive juice, after the digestion in the water bath, a large part of the digestive juice is still unused, resulting in waste of digestive juice, and the utilization rate of digestive juice is very 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
  • Rat II type primary alveolar epithelial cell extraction method
  • Rat II type primary alveolar epithelial cell extraction method
  • Rat II type primary alveolar epithelial cell extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Equipment needed: healthy rats, large petri dishes, intravenous infusion needles, catheters (intravenous infusion needles), black thread, 15ml pancreatin, 15ml type I collagenase, syringes, centrifuge tubes.

[0022] Steps:

[0023] ①Bind the intravenous infusion needle and the trachea: dissect the bronchi and the two lungs connected to it from the rat, flush the residual blood, remove the necrotic tissue, place it in a large petri dish, and place the intravenous infusion needle 1 cm from the tip with forceps Break it and retain the 1cm needle (to prevent the sharp needle tip from puncturing the trachea), insert the intravenous needle into the trachea of ​​the lung, fix the trachea and the needle with a hemostatic forceps, and then bind the intravenous needle and trachea with a black thread ( 3 turns on the top and bottom) (such figure 1 Shown);

[0024] ②Quickly inject the digestive juice: Pour 15ml 0.25% pancreatin and 15ml 0.1% type I collagenase into a 50ml centrifuge tub...

Embodiment 2

[0031] Equipment needed: healthy rats, large petri dishes, intravenous infusion needles, catheters (intravenous infusion needles), black thread, 15ml pancreatin, 15ml type I collagenase, syringes, centrifuge tubes.

[0032] Steps:

[0033] ①Bind the intravenous infusion needle and the trachea: dissect the bronchi and the two lungs connected to it from the rat, flush the residual blood, remove the necrotic tissue, place it in a large petri dish, and place the intravenous infusion needle 1 cm from the tip with forceps Break and keep the 1.2cm needle (to prevent the sharp needle from puncturing the trachea), insert the intravenous needle into the trachea of ​​the lung, fix the trachea and the needle with a hemostatic forceps, and then bind the intravenous needle and trachea with a black thread (3 turns on each side) (e.g. figure 1 Shown);

[0034] ②Quickly inject the digestive juice: Pour 15ml 0.25% pancreatin and 15ml 0.1% type I collagenase into a 50ml centrifuge tube and mix evenly ...

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 relates to the technical field of biological cell culture, and specifically relates to a rat II type primary alveolar epithelial cell extraction method, which comprises following steps:(1) binding an intravenous infusion needle and a tracheae; (2) rapidly injecting a digestion solution; (3) putting lung tissues into a centrifuge tube; (4) absorbing the digestion solution; (5) slowlyinjecting the digestion solution; (6) reusing the digestion solution; and (7) performing cyclic operation. The primarily added digestion solution is cyclically used, the digestion solution is repeatedly absorbed and injected into a lung to carry out digestion, the amount of digestion solution that is injected into the lung is basically equal to the amount of digestion solution that exudes throughthe lung so as to ensure that the pulmonary alveoli is filled with the digestion solution; the digestion becomes stable, continuous, rapid, and more complete; the total digestion time is largely shortened, the digestion efficiency and utilization rate of the digestion solution are effectively improved; and the yield of II type primary alveolar epithelial cells is increased.

Description

Technical field [0001] The invention relates to the technical field of biological cell culture, in particular to a method for primary extraction of rat type II alveolar epithelial cells. Background technique [0002] Rat type Ⅱ alveolar epithelial cells are isolated from lung tissue; type Ⅱ alveolar epithelial cells have the potential to divide, proliferate and differentiate into type Ⅰ alveolar cells, so they have the effect of repairing damaged epithelium. At present, the method of culturing rat type II alveolar epithelial cells is mainly enzymatic digestion. The animal source is mainly the most common, practical and convenient experimental animal in the field of life science research—rats. Rats are extracted in the prior art. The method for type Ⅱ alveolar epithelial cells is to first inject a mixture of pancreatin and type Ⅰ collagenase (digestive juice) into the rat’s lungs, place the lungs in a centrifuge tube, and then place the centrifuge tube containing the lungs Digest...

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): C12N5/071
CPCC12N5/0688C12N2509/00
Inventor 陈博文陈淼蒲清湖王洪亮刘国跃
Owner AFFILIATED HOSPITAL OF ZUNYI MEDICAL COLLEGE
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