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Stem cell factor alters membrane potential of purified peritoneal mast cells in culture
1Department of Medicine, School of Medicine, University of California, Los Angeles 90095, USA.
Abstract:
The membrane potential (E(m)) was used as an indicator to evaluate the effect of stem cell factor (SCF) on the membrane integrity of peritoneal mast cells (PMCs). PMCs were harvested from the peritoneal lavage of Sprague-Dawley rats, purified more than 95% and cultured with or without the presence of SCF (2 x 10(-8) M). E(m) values were measured with conventional intracellular recording techniques. Results from day 1 to day 4 in culture were compared. Significant differences in average E(m) (aE(m)) (P < 0.01, analysis of variance) were seen on days 3 and 4 (means +/- SE in millivolts): -67.4 +/- 8.0 and -59.4 +/- 4.8 with SCF vs. -24.8 +/- 7.9 and -7.6 +/- 3.9 without SCF, respectively. Moreover, after culture with SCF for >1 wk, the aE(m) values of purified PMCs had a tendency to reach plateau values similar to that of unpurified PMCs on day 1 (at -20 mV). The morphological appearances of PMCs can be correlated with the results of aE(m) measurements. PMCs with a smooth spherical shape and highly refractive appearance, and better tolerance to electrode impalement, showed E(m) with greater negative values and lesser fluctuations. These results indicate that SCF can maintain the membrane properties and viability of purified PMCs in a long-term culture.
Insights
Stem cell factor (SCF) maintains the membrane integrity and viability of purified peritoneal mast cells (PMCs) in long-term cultures. SCF treatment resulted in significantly more negative membrane potential values compared to untreated cells.
Area of Science:
- Immunology
- Cell Biology
- Physiology
Background:
- Peritoneal mast cells (PMCs) are crucial immune cells.
- Maintaining PMC membrane integrity during in vitro culture is challenging.
- Stem cell factor (SCF) is known to influence mast cell development and function.
Purpose of the Study:
- To investigate the effect of SCF on the membrane potential (E(m)) and integrity of purified PMCs in culture.
- To assess the long-term viability of PMCs cultured with SCF.
Main Methods:
- Purified PMCs (>95%) from Sprague-Dawley rats were cultured with or without SCF (2 x 10(-8) M).
- Membrane potential (E(m)) was measured using conventional intracellular recording techniques.
- Morphological appearances were correlated with E(m) measurements.
Main Results:
- SCF-treated PMCs exhibited significantly more negative average E(m) values on days 3 and 4 compared to controls (P < 0.01).
- SCF treatment promoted E(m) values in purified PMCs that approached those of unpurified PMCs after 1 week.
- Improved membrane integrity, indicated by greater negative E(m) and lesser fluctuations, correlated with a smooth spherical morphology.
Conclusions:
- SCF effectively maintains the membrane properties and viability of purified PMCs during long-term culture.
- SCF is essential for preserving the physiological characteristics of PMCs in vitro.
- These findings support the use of SCF for long-term studies involving purified PMCs.