Isolation of Ca2+ Tolerant Cardiomyocytes from Aadult Rats for Patch Clamp Studies
XU Hua-e
Tao Jin
CHEN Jie
Wang Hong-yi
LI Sheng-nan
Abstract：Objective: To investigate the factors affecting the viability and Ca2+ tolerance of isolated rats' cardiac myocytes for patch clamp research. Methods: Hearts were firstly perfused by the Langendorff perfusion apparatus with normal Tyrode' s solution, then with Ca2+-free Tyrode's solution and subsequently with Iow Ca2+ enzyme solution containing collagenase 0.1-0.2 g/L. All the solutions were saturated with oxygen and the perfusion temperature was kept at 37 ℃. Finally hearts were washed by Ca2+-free Tyrode' s solution, after which the ventricles were minced into small pieces in KB solution, dispersed and filtered. The isolated myocytes were stored in KB solution at room temperature for 1 h and recovered to normal calcium concentration before patch clamp experiments. Results: When all the factors such as water, enzyme, Ca2+ ,pH, and oxygen were well controlled, the well constructed and rod-like cardiac myocytes with a yielding rate of 30%-50% came out. Conclusion: All the factors should be well controlled, which ensured the isolated cells Ca2+ tolerant and appropriate for patch clamp experiments.
Keywords：isolationcardiac myocytepatch-clampTyrode
Publication Date：2004-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 4-6 )