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Highly insulin-responsive isolated rat heart muscle cells yielded by a modified isolation method
Y Fischer1, H Rose, H Kammermeier
1Institute of Physiology, Medical Faculty, RWTH Aachen, F.R.G.
Life Sciences
|January 1, 1991
Summary
Researchers developed a modified method for isolating rat cardiomyocytes, achieving a low basal glucose uptake rate. This new method creates a highly responsive model for studying glucose transport stimulators like insulin.
Area of Science:
- Cardiology
- Cell Biology
- Metabolism
Background:
- Isolated adipocytes and cardiac myocytes often show unspecific stimulation during handling, affecting glucose transport.
- Previous methods resulted in higher basal hexose uptake rates in isolated cardiomyocytes compared to physiological levels.
Purpose of the Study:
- To develop a modified isolation procedure for rat cardiomyocytes yielding a low basal glucose uptake rate.
- To establish a more physiologically relevant and responsive model for studying glucose transport mechanisms.
Main Methods:
- Modified isolation procedure for rat cardiomyocytes.
- Measurement of basal and insulin-stimulated hexose (2-deoxy-D-glucose and 3-O-methyl-D-glucose) uptake.
- Assessment of stimulatory effects of sulfhydryl reagents.
Main Results:
- The modified procedure yielded cardiomyocytes with a significantly low basal hexose uptake rate (approx. 3 pmol*s-1*mg protein-1).
- Insulin (10 nM) demonstrated a marked stimulatory effect (8- to 20-fold) on glucose uptake, exceeding that typically observed in isolated cells.
- The cells responded to other stimulators, such as phenylarsine oxide, indicating preserved responsiveness.
Conclusions:
- The modified method provides rat cardiomyocytes with basal glucose transport rates close to physiological values.
- These isolated cardiomyocytes represent a robust and sensitive model for investigating glucose transport regulation and the effects of various stimulators.