Hi there,

  I have two questions for the isolation of murine single lung cells:

  1. We use collagenase I for lung tissue digestion for 1h at 37 ℃ after cutting the lung tissues into small pieces. After the digestion, the lung tissues are still being pieces and we smash the tissues against 70 um cell strainer to get the single cells. Is it normal that the lung tissues are still pieces after the digestion?

  2. Sometimes we can see that the isolated cells clump together and we lost some cells. Is the 2mM EDTA helpful to prevent the situation or are there any other methods to make it better?

Many thanks for your reply.