Aim:
I am isolating adult cardiomyocytes from mice injected by our therapeutic oligo and my aim is to measure its uptake.
Method:
I am able to isolate viable cardiomyocytes using the classic Langendorff setup with Collagenase + Protease digestion. After collecting and purifying the viable cells (i.e. "rod shaped" cardiomyocytes) I then sonicate them to lyse and release all the intracellular oligo, which is then measured using our published ELISA method.
Concern:
One of my biggest concern is the intracellular oligos "leaking" out of the cell during the 2hr isolation process. Therefore I wonder if there is a way I can "encapsulate" the cells to prevent possible leakage before eventually lysing them?
One step at a time, Mr. Burki. You fist need solid evidence of the intracellular oligos actually leaking out the cell before you think of a way to prevent it. Label your oligos and then separate the cells from extracellular solution to see if there is actually a leakage. If this labeling experiment suggests there is leakage, then you can think of ways to prevent it.
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