Method and application of large proliferation of newborn mouse skin terminally isolated cells

Abstract

The invention belongs to the technical field of animal cell culture, and relates to a method and application of large proliferation of newborn mouse skin terminally isolated cells. The method comprises the following steps that a newborn mouse skin sample is pretreated; an enzyme digestive liquid is prepared; the sample is treated by the enzyme digestive liquid, and terminally isolated cell suspension liquid is obtained; primary cells of the newborn mouse skin terminally isolated cells are prepared; passage and expansion culture is conducted on the primary cells; passage cells are prepared; andthe skin terminally isolated cells are screened and purified by a c-kit+/CD34+ flow cytometry, the cell sap is resuspended and subjected to expansion culture, and the skin terminally isolated cells with the high purity are prepared. The method adopts collagenase II and trypsin with the concentration being both 0.05% to separate and culture the skin terminally isolated cells by combining with a digestion method, the number of the obtained primary terminally isolated cells is large, the purity is high, the activity is good, the cells can be rapidly and largely proliferated after being screenedby the flow cytometry, and the proliferated cells are stable in morphology and high in activity, and can be widely applied to morphological detection and function research of the follow-up related terminally isolated cells.

Description

technical field [0001] The invention relates to the technical field of animal cell culture, in particular to a method for amplifying a large number of neonatal rat skin terminal septal cells and its application. Background technique [0002] Telocytes (TCs), a newly discovered and named interstitial cell, are widely distributed in skin, heart, kidney, liver, gastrointestinal tract, bladder, parotid gland, placenta, lung, bronchi and brain, etc. in an organization. Septal terminal cells have typical morphological features: small cell body, one or more elongated processes (telopodes, TPs) of different lengths protruding from the cell body, with constriction (podomer) and enlargement (podom) on the protrusions Arranged alternately, showing a bead-like structure, forming a complex three-dimensional network with surrounding mesenchymal cells. [0003] For the identification of terminal septal cells, observation of tissue-level morphological characteristics by transmission elect...

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For example, in the esophagus, type II collagenase and deoxyribonuclease are configured as digestive fluid, the centrifugation rate of cells is 2000rpm, the cell culture differential attachment time is 60-90 min, and the cell culture medium is high-sugar DMEM medium; for Pr

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