Experiments in the preparation of extracellular matrix can be used to (1) study the constituents of the extracellular matrix as well as its physiological characteristics, and (2) observe the specific roles it plays in different tissues.

Operation method

Preparation of extracellular matrix (ECM)

Principle

Remove the substrate with a decontaminant to form a postcellular confluent monolayer (postoonfluent monolayer), wash the culture flask or dish, and then culture the desired cells on the residual substrate.

Materials and Instruments

3T3 mouse fibroblasts MCR-5 human fibroblasts CPAE bovine pulmonary artery endothelial cells or any other cell line suitable for the production of extracellular matrix.
Sterile ultrapure water (UPW), sterilized l% TritonX-100
Culture flasks or dishes

Move

makings

3T3 mouse fibroblasts, MCR-5 human fibroblasts, CPAE bovine pulmonary artery endothelial cells, or any other cell line suitable for extracellular matrix production.

Sterile ultrapure water (UPW).

Sterilized l% TritonX I 100 and UPW.

Procedure

1. Cultivate stroma-producing cells until the culture vessel is full.

2. After 3-5 days to reach confluence, remove the medium and add an equal volume of sterile 1% TritonX-100 prepared in UPW to the monolayer.

3. Incubate at 37℃ for 30min.

4. Remove the TritonX-100 solution and rinse three times with an equal volume of UPW.

5. The flasks or petri dishes can be used directly or left at 4℃ and used within 3 weeks.

Caveat

The flasks or petri dishes can be used directly or left at 4°C and used up within 3 weeks.

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