What plating densities are recommended, in general, for cardiomyocytes? Is this the same for dissociation and patch clamp applications?

What plating densities are recommended, in general, for cardiomyocytes? Is this the same for dissociation and patch clamp applications?


If you opened this Solution from a search, your last search results are still available in a separate tab. Click the button below to begin a new search.

SEARCH ALL SOLUTIONS

Still Have Questions?
If you did not find an answer to your question, please contact us to speak with an expert:

(877) 320-6688 (toll-free)
+1 (608) 310-5100
or submit a support request

The optimal plating density for iCell Cardiomyocytes can be assay dependent and should be determined empirically based on the intended use. However, a density of ~63,000 plated cardiomyocytes/cm2 provides a beating syncytium and is the recommended starting density for most cell-based assays. See the table below for the recommended number of plated cells for different culture formats.

Culture Format Plated Cells Volume
T-25 Flask 1,200,000 – 1,600,000 8 ml
6-well Plate 600,000 2 – 3 ml
12-well Plate 240,000 1 – 1.2 ml
24-well Plate 120,000 500 – 600 µl
96-well Plate 20,000 100 µl
384-well Plate 5,000 25 – 40 µl
1536-well Plate 2,000 8 – 10 µl

For dissociation applications, we recommend plating at a higher plating density of 750,000 viable cells/well in a 6-well plate according to our Application Protocol.

To obtain single cells that are suitable for manual patch clamp, we recommend plating 40,000 – 80,000 viable cells/well in 2 ml total volume in a 12-well plate containing a coverslip according to our Application Protocol.

References:

  1. iCell Cardiomyocytes User’s Guide
  2. Application Protocol: Dissociating Cardiomyocytes by Enzymatic Treatment 
  3. Application Protocol: Measuring Cardiac Electrical Activity: Manual Perforated Patch Clamp