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...

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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

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