Search of Neuroprotective Polyphenols Using the "Overlay" Isolation Method

Overview

Journal Molecules

Publisher MDPI

Specialty Biology

Date 2018 Jul 26

PMID 30042342

Citations 10

Authors

Hiroshi Sakagami

Haixia Shi

Kenjiro Bandow

Mineko Tomomura

Akito Tomomura

Misaki Horiuchi

Tomohiro Fujisawa

Takaaki Oizumi

Affiliations

Soon will be listed here.

Abstract

Previous studies of the neuroprotective activity of polyphenols have used ununiform culture systems, making it difficult to compare their neuroprotective potency. We have established a new and simple method for preparing differentiated PC12 cells by removing the toxic coating step. Cells were induced to differentiate with the nerve growth factor (NGF) in a serum-free medium, without a medium change, but with a one-time overlay supplementation of NGF. The optimal inoculation density of the cells was 6⁻12 × 10³ cells/cm², and the presence of serum inhibited the differentiation. Neuroprotective activity could be quantified by the specific index (SI) value, that is, the ratio of the 50% cytotoxic concentration to the 50% effective concentration. Alkaline extract from the leaves of Rehder (SE), having had hormetic growth stimulation, showed the highest SI value, followed by epigallocatechin gallate. The SI value of curcumin and resveratrol was much lower. This simple overly method, that can prepare massive differentiated neuronal cells, may be applicable for the study of the differentiation-associated changes in intracellular metabolites, and the interaction between neuronal cells and physiological factors.

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