Elimination of platelets from mononuclear cell preparations using heat-killed yeast
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Published on: March 27, 2014
Abstract:
A new method for reducing platelet contamination in mononuclear cell preparations using heat killed yeast is presented. Yeast cells are added to the buffy coat of peripheral blood, incubated at 37 degrees C for 1 hour, layered over a Ficoll-Hypaque gradient, and centrifuged at 700 g for 30 minutes at room temperature. The cells at the interface are collected and evaluated for platelet count, lymphocyte surface markers, and mononuclear cell recovery. With increasing yeast concentration, the platelet count decreases to 93% over control values. No detectable lymphocyte subset is removed by this method. Cell recovery varies, but seems to decrease with the addition of higher concentrations of yeast cells. Normal plasma is required for the successful removal of platelets. Heat-inactivated plasma significantly reduces the efficiency of platelet removal.
Insights
This study introduces a novel method using heat-killed yeast to reduce platelet contamination in mononuclear cell preparations. The technique effectively lowers platelet counts without affecting lymphocyte subsets, crucial for cell isolation accuracy.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Mononuclear cell preparations are vital for various research and clinical applications.
- Platelet contamination can interfere with downstream analyses and cell function.
- Existing methods for platelet reduction may impact mononuclear cell viability or subsets.
Purpose of the Study:
- To develop and evaluate a new method for reducing platelet contamination in mononuclear cell preparations.
- To assess the impact of heat-killed yeast on platelet counts and mononuclear cell recovery.
- To determine the influence of plasma conditions on the efficacy of platelet removal.
Main Methods:
- Peripheral blood buffy coats were treated with heat-killed yeast.
- Cells were incubated, layered over Ficoll-Hypaque, and centrifuged.
- Platelet counts, lymphocyte surface markers, and cell recovery were analyzed.
Main Results:
- Increasing yeast concentration reduced platelet counts by up to 93% compared to controls.
- No significant depletion of lymphocyte subsets was observed.
- Cell recovery decreased with higher yeast concentrations; normal plasma was essential for optimal results.
Conclusions:
- Heat-killed yeast provides an effective method for reducing platelet contamination in mononuclear cell preparations.
- The method preserves lymphocyte subsets, offering a valuable tool for cell isolation.
- Optimizing yeast concentration and maintaining normal plasma are critical for successful application.
