Stimulating Nerve Stem Cell Differentiation Using Cold Atmospheric Plasma Treatment

Abstract

Source: Xiong, Z., et al. Nerve stem cell differentiation by a one-step cold atmospheric plasma treatment in vitro. J. Vis. Exp. (2019).

This video presents a method for triggering differentiation in neural stem cells by utilizing cold atmospheric plasma. This generates UV radiation and reactive molecules to stimulate nerve cells to produce projections, indicating neuronal cell differentiation.

Protocol

1. Cell Cultures and Predifferentiation

1. Neural stem cell culture and predifferentiation
   1. Prepare poly-D-lysine-coated coverslips. Put a sterile coverslip (20 mm in diameter) into a 12-well plate. Coat the cover glass with poly-D-lysine, 0.1% w/v, in water for better cell adhesion on the coverslips by following the next steps.
      ​NOTE: Optimal conditions must be determined for each cell line and application.
      1. Aseptically coat the surface of the coverslip with poly-D-lysine, 0.1% w/v, in water. Rock gently to ensure an even coating of the coverslip surface.
      2. After culturing overnight (~12 h) at ....

Results

Figure 1: Schematic of the experimental set-up. The plasma jet device is connected to the output wire of the high-voltage power supply. The high-voltage probe with oscilloscope is used to detect the output voltage. When the working gases are flowing th.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Coverslip	NEST	801008
Poly-D-lysine	Beyotime	P0128
DMEM medium	HyClone	SH30022.01B	stored at 4 °C
DMEM/F12 medium	HyClone	SH30023.01B	stored at 4 °C
N2 supplement	Gibco	17502048	stored at -20 °C and protect from light
B27 supplement	Gibco	17504044	stored at -20 °C and protect from light
Fetal bovine serum	HyClone	SH30084.03	stored at -20 °C, avoid repeated freezing and thawing
Donor Horse serum	HyClone	SH30074.03	tored at -20 °C, avoid repeated freezing and thawing
Penicillin/Streptomycin	HyClone	SV30010	stored at 4 °C
Trypsin	HyClone	25300054	stored at 4 °C
PBS solution	HyClone	SH30256.01B	stored at 4 °C
4% paraformaldehyde	Beyotime	P0098	stored at -20 °C
TritonX-100	Sigma	T8787
Light microscope	Nanjing Jiangnan Novel Optics Company	XD-202
High – voltage Power Amplifier	Directed Energy	PVX-4110
DC power supply	Spellman	SL1200
Function Generator	Aligent	33521A
Oscilloscope	Tektronix	DPO3034
High voltage probe	Tektronix	P6015A
12-well plate	corning	3512
25 cm2 flask	corning	430639
Helium gas
Oxygen Gas
Plasma jet nozzel
CO2 Incubator
Silicone pipe			inner diameter of 12 mm
Murine neural stem cell C17.2 (C17.2-NSC)