I was coculturing immortalized bone-marrow derived macropahges with Bifidobacteria. Any heads up on how to lyse these cells for plating the lysates to check bacterial persistence within these cells? Thanks.
Hi Sreeram, I work with primary human alveolar macrophages and monocyte derived macrophages infected with M. tuberculosis. I lyse the infected macrophages with 0.1% SDS for 5 minutes and then stop the lysis with 20% BSA. That is sufficient to recover the intracellular bacteria. I make serial dilutions of the lysate and plate them in triplicates. Everything works fine.