Enhanced nerve growth factor efficiency in neural cell culture by immobilization on the culture substrate.
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 100 ng of NGF to cultures daily, and compared them to the effects of immobilizing 5, 10, or 100 ng of NGF on culture substrates in a single dose. Cultures with 10 or 5 ng NGF added daily showed dramatically decreased cell viability, mitochondrial metabolic activity, and neuronal differentiation compared to cultures with 100 ng NGF added daily, while also exhibiting increased apoptosis. In contrast, a single dose of 100 ng immobilized NGF yielded results similar to 100 ng NGF added daily (total: 300 ng over 3 days), and 10 or 5 ng immobilized NGF showed far better results than 10 or 5 ng NGF added daily. These results demonstrate that NGF immobilization can dramatically reduce the amount of NGF required in neuronal cell culture.
Bhang
Suk Ho
SH
Department of Bioengineering, Hanyang University, Sungdong-gu, Seoul, Republic of Korea.
Lee
Tae-Jin
TJ
Yang
Hee Seok
HS
La
Wan-Geun
WG
Han
Ah Mi
AM
Kwon
Yun Hee Kim
YH
Kim
Byung-Soo
BS
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D055767:Immobilized Proteins / Q000494:pharmacology*; D020932:Nerve Growth Factor / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D016716:PC12 Cells; D051381:Rats
D055767:Immobilized Proteins; D020932:Nerve Growth Factor