Hi Natalie, I would recommend that you have a look at the recent review by my former colleagues (Voigt et al. J Mol Cell Cardiol. 2015 Sep;86:187-98) that addresses this in detail! Good luck.

Jordi

Hello Natalie,

Hello!

If you are interested in isolation of cardiomyocytes from humans, please follow the chunk procedure. Here is the method. You can find it in the recent paper Glukhov, Balycheva et al. Circ. 2015 "Direct Evidence for Microdomain-Specific Localization and Remodeling of Functional L-Type Calcium Channels in Rat and Human Atrial Myocytes". I used this approach during 3 years and it gave me nice and healthy myocytes which were totally ok for patch-clamp (even 2 days in culture), staining, contraction etc. You can use this protocol for both atrial and ventricular cells. In case of fibroblast just simply collect supernatant after each centrifuging.

Briefly, individual specimens were transferred to ice-cold calcium free Krebs-Ringer saline solution consisting of (in g/L): 7.012 NaCl, 0.402 KCl, 1.332 MgSO

4, 0.55 Pyruvate, 3.603 Glucose, 2.502 Taurine, 2.383 HEPES, 1.286 Nitrillotriacetic Acid; pH = 6.96. Connective and adipose tissue were removed and approximately 500mg of myocardial tissue was minced with razor blades in small cubes (approx. 1-2 mm 3). Then, the tissue pieces were washed with fresh Ca2+-free Krebs-Ringer solution 3 times for 3 min each at 37C. After wash, cardiac tissue was incubated for 25 min in 10ml of Krebs-Ringer solution containing (in g/L): NaCl 7.012, KCl 0.402, MgSO4 1.332, Pyruvate 0.55, Glucose 3.603, Taurine 2.502, HEPES 2.383; pH = 7.4, supplemented with 200 nM CaCl2 and Proteinase type XXIV (0.36mg/ml; Sigma-Aldrich) under gentle agitation. The partially digested tissue was transferred to 10ml of Krebs-Ringer saline supplemented with collagenase type XIV (1mg/ml Sigma-Aldrich). The tissue was incubated thrice with this solution for 10 min each at 37C with gentle agitation. Usually, cardiomyocytes were visible by phase contrast light microscopy after the first incubation step, with the biggest amount of cells after the second incubation step. After each incubation step, the supernatants were transferred to a tube and centrifuged at 600 rpm for 3 min. The pellets were re-suspended in 2-3 mL of Krebs-Ringer solution.

Good luck!

Hello

Look at links here will help you 

Good Luck

https://www.wpi.edu/Pubs/E-project/Available/E-project-042612-144641/unrestricted/Automated_Cardiomyocyte_Isolation_Report.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3164875/

http://cardiovascres.oxfordjournals.org/content/cardiovascres/58/2/423.full.pdf

Creative Bioarray provides high-quality primary cells.

Maybe you can scan via https://www.creative-bioarray.com/Human-Cardiomyocytes-CSC-C2847-item-39324.htm

https://www.creative-bioarray.com/QualiCell®-Human-Cardiomyocytes-transdiff-from-Human-Stem-Cell-XLC450-CSC-C00280-item-2836.htm

Good luck :)