This study explores the differentiation of nerve stem cells using cold plasma treatment. The process involves culturing stem cells on an extracellular matrix and applying plasma jets to activate differentiation.
Begin with nerve stem cells cultured on an extracellular matrix-coated coverslip in a suitable differentiation medium.
Take a syringe containing a cold plasma, which is a state of matter containing various reactive components, including UV radiation and charged particles.
Position the nozzle of the plasma syringe at a specific height above the nerve cell culture.
Start the plasma jet.
The plasma interacts with the nerve stem cells, activating them.
Remove the old culture medium. Add a fresh culture medium and incubate.
Over time, the activated nerve stem cells produce projections.
Discard the medium.
Under an inverted phase contrast light microscope, the presence of elongated projections on the cells, resembling those found in a nerve cell, indicates differentiation.
To plasma-treat treat the neural stem cells, set the distance between the syringe nozzle and the first well of cells to 15 millimeters. Replace the supernatants in the neural stem cell cultures with 800 microliters of fresh differentiation medium, and place the plate under the plasma jets.
Making sure that the syringe nozzle is fixed over the center of each well, create three wells with plasma for 60 seconds per treatment, and three wells with 1% helium and oxygen gas for 60 seconds per treatment, leaving three wells untreated as controls. Then, replace the supernatants with one milliliter of fresh differentiation medium, and return the cells to the incubator for six days. Check the differentiation status of the different groups daily under an inverted phase contrast light microscope.
After the treatment, allow the cells to fully equilibrate in the conditioned medium for about one hour before replacing the supernatant with fresh differentiation medium.
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