Enhanced nerve growth factor efficiency in neural cell culture by immobilization on the culture substrate

• Published in: Biochemical and biophysical research communications Vol. 382; no. 2; pp. 315 - 320
• Main Authors: Bhang, Suk Ho, Lee, Tae-Jin, Yang, Hee Seok, La, Wan-Geun, Han, Ah Mi, Kwon, Yun Hee Kim, Kim, Byung-Soo
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2009
• Subjects: Animals; Cell Culture Techniques; Cell Differentiation; Immobilization; Immobilized Proteins - pharmacology; Nerve growth factor; Nerve Growth Factor - pharmacology; Neuronal differentiation; Neurons - cytology; Neurons - drug effects; PC12 Cells; Pheochromocytoma cell; Rats; Immobilization; Neuronal differentiation; Nerve growth factor; Pheochromocytoma cell
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2009.03.016

Nerve growth factor (NGF) immobilization on a culture substrate may dramatically reduce the amount of NGF required for pheochromocytoma (PC12) cell culture. Coverslips on which NGF had been immobilized, or with NGF added to the culture medium daily, were used to culture PC12 cells. We examined the effects of adding 5, 10, or 100ng of NGF to cultures daily, and compared them to the effects of immobilizing 5, 10, or 100ng of NGF on culture substrates in a single dose. Cultures with 10 or 5ng NGF added daily showed dramatically decreased cell viability, mitochondrial metabolic activity, and neuronal differentiation compared to cultures with 100ng NGF added daily, while also exhibiting increased apoptosis. In contrast, a single dose of 100ng immobilized NGF yielded results similar to 100ng NGF added daily (total: 300ng over 3days), and 10 or 5ng immobilized NGF showed far better results than 10 or 5ng NGF added daily. These results demonstrate that NGF immobilization can dramatically reduce the amount of NGF required in neuronal cell culture.