A simple, rapid and cost-effective method to isolate the huge quantity of human mononuclear cells from minute quantities of blood as well as buffy coat for different immunological and magnetic beads applications
Description
Aim: Today the isolation of mononuclear cells (MNC) is being done with density gradient methods, which are cumbersome and time consuming, but need minimum 10 ml of buffy coat or blood to be performed, as this method is unable to isolate the MNC from minute volume like 100 µl. Therefore, we decided to develop a simple, rapid and cost-effective method to isolate the MNC from minute volume of blood and buffy coat.
Methods. MNC isolator was used to isolate the MNC from 100 as well as 500 µl of blood as well as buffy coat just mixing and centrifugation. Isolated cells were cultured and magnetic beads were used to isolate the CD4 and CD45 specific cells.
Results: In 15 experiments, MNC isolator were used successful to isolate MNC from minute volumes of blood and buffy coat. The cell cultured were generated and they were maintained up to 3 years without any contamination and the cells remain healthy. Successful isolation of CD4 and CD45 cells with magnetic beads war carried. The cells were frozen and thawed.
Conclusions: In this work, we developed first time a simple, cost effective and robust method to isolate MNC from minute volume of buffy coat and blood for immunological applications and magnetic beads cell isolation. The method is so simple that it can be used in any cell culture laboratory in the world. This method should open new opportunities to develop new kinds of antibodies, to isolate the different cell populations with magnetic beads as well as to get large number of T-cells for development of immunotherapies like CAR T-cell therapies.
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27.09_isolate the mononuclear cells from small quantity.pdf
Additional details
References
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