1, collagenase Ⅰ used in epithelium, lungs, separation of fat and adrenal tissue cells.It is used in digesting the connective tissue to make it a single cell and in the separation of mammalian cells.

2, collagenase Ⅱ applies to the liver, bone, thyroid, heart and salivary glands.

3, collagenase Ⅳ contain at least seven kinds of protease, the molecular weight range from 68-130 kd.It can digest a variety of tissues.

4, collagenase Ⅴ contain at least seven kinds of protease, the molecular weight range from 68-130 kd.It can be used to separate small island tissues of the pancreas, separating connective tissue into individual cells.

Rinse and trim the tissue into 1 to 2mm3 pieces.

Place the tissue mass into a centrifuge tube (triangle flask) and add 30 to 50 times the volume of collagenase solution. Tighten the lid (seal the flask).

Place the flask in a water bath at 37℃ or a incubator at 37℃, and shake it every 3min. It is better to place it in a constant temperature vibrating water bath at 37℃.The digestion time is closely related to the type of tissue. For some tumor tissues or other relatively dense connective tissues, the digestion time is 4~48h. For easily digested tissues, shock digestion at 37℃ for 15~45min can also be used, depending on the specific situation.If the tissue mass has dispersed and lost the shape of the mass, the cell mass or individual cell can be considered fully digested when shaken.After collagenase digestion of epithelial tissue, there may still be some cells that are not fully dispersed due to the resistance of epithelial cells to this enzyme. However, epithelial cells in groups are more likely to grow than isolated epithelial cells.

The digestive juice (sometimes containing individual tissue blocks and tissue fragments that are not fully digested should be filtered with 100 mesh stainless steel mesh) was collected, centrifuged at 1000r/min for 5min, and then the supernatant was removed.

The material came from the Internet.

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