Growth and Differentiation of Magnetic Nano Particle-Loaded Neurons on Magnetic Platforms

Abstract

Source: Indech, G., et al. Fabrication of Magnetic Platforms for Micron-Scale Organization of Interconnected Neurons. J. Vis. Exp. (2021).

This video demonstrates the uptake of fluorescent magnetic nanoparticles (MNPs) by the neuronal cells via endocytosis. Growing these MNP-loaded neurons on the magnetically patterned substrate enables the attachment of the cells. The addition of a growth factor containing medium aids in the growth of neuron extensions, while the magnetic field of the substrate directs the orientation of the extension.

Protocol

1. Cellular magnetic nanoparticle (MNP) uptake

1. Prepare basic growth medium for PC12 cell culture by adding 10% horse serum (HS), 5% fetal bovine serum (FBS), 1% L-glutamine, 1% penicillin/streptomycin, and 0.2% amphotericin to Roswell Park Medical Institute (RPMI) medium, and filter using a 0.22 µm nylon filter.
2. Add 1% horse serum (HS), 1% L-glutamine, 1% penicillin/streptomycin, and 0.2% amphotericin to RPMI medium to prepare PC12 differentiation medium and filter using a 0.22 µm nylon filter.
3. Grow cells in a non-treated culture flask with 10 mL of basic growth medium; add 10 mL of basic growth medium to the ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
6-well cell culture plate	FALCON	353846
96-well cell culture plate	SPL life sciences	30096
Amphotericin B solution	Biological Industries	03-028-1B
Bovine serum albumin (BSA)	Biological Industries	03-010-1B
Cell and Tissue cultur flask	Biofil	TCF002250	75.0 cm^2 250 mL Vent cap, Non-treated
Cell culture dish	Greiner Bio-One	627-160	35 mm
Centrifuge tube	Biofil	CFT021500	50 mL
Collagen type I	Corning Inc.	354236	Rat Tail, concentration range 3-4 mg/mL
Disposable needle	KDL		23 G
Disposable syringe	Medispo	1160227640	10 mL
Donor horse serum	Biological Industries	04-124-1A
Ethanol 70%	ROMICAL LTD	19-009102-80
Ethanol absolute (Dehydrated)	Biolab-chemicals	52505
Fetal bovine serum (FBS)	Biological Industries	04-127-1A
Fresh murine β-NGF	Peprotech	450-34
GMW C-frame electromagnet .	Buckley systems LTD		3470, 45 mm
L-glutamine	Biological Industries	03-020-1B
PBS 10x	hylabs	BP507/1LD
PC12 cell line	ATCC	CRL-1721
Penicillin-streptomycin nystatin solution	Biological Industries	03-032-1B
RPMI 1640 with l-glutamine	Biological Industries	01-100-1A