Rat II type primary alveolar epithelial cell extraction method

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...

Examples

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 ...

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...

Examples

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 ...

Images