Bilirubin Cytotoxicity in Primary Mouse Cerebral Cortical Cell Culture.

Choi,  Chang Won; Min,  Ki Woon; Kim,  Mi Nyeu; Hwang,  Jong Hee; Shim,  Jae Won; Koh,  Sun Young; Chang,  Yun Sil; Park,  Won Soon

Original Article

Journal of the Korean Society of Neonatology 2003;10(2):241-247.
Published online November 1, 2003.

Bilirubin Cytotoxicity in Primary Mouse Cerebral Cortical Cell Culture.

Chang Won Choi, Ki Woon Min, Mi Nyeu Kim, Jong Hee Hwang, Jae Won Shim, Sun Young Koh, Yun Sil Chang, Won Soon Park

¹Department of Pediatrics, Samsung Seoul Hospital, College of Medicine, Sungkyunkwan University, Seoul, Korea. wspark@smc.samsung.co.kr
²Department of Pediatrics, Kangbuk Samsung Hospital, College of Medicine, Sungkyunkwan University, Seoul, Korea.
³Department of Pediatrics, Samsung Cheil Hospital, College of Medicine, Sungkyunkwan University, Seoul, Korea.

일차배양된 마우스의 대뇌피질세포에서 빌리루빈의 세포독성에 대한 연구

^, ^, ^, ^, ^, ^, ^,

^¹
^²
^³

Abstract

PURPOSE
We sought to quantitate the cytotoxicity of bilirubin for neuronal cells at various concentrations of bilirubin and at various [bilirubin]/[albumin] ratio.
METHODS
Mouse cerebral cortical cells were obtained from 15 day-old mouse fetal cerebral cortex primary culture. Cerebral cortical cells were exposed to medium containing various concentrations of bilirubin and [bilirubin]/[albumin] ratios for 4 hours. Then, the bilirubin cytotoxicity for cerebral cortical cells was quantitated by the activity of lactate dehydrogenase (LDH) released from cerebral cortical cells into the culture media and the viability of cerebral cortical cells was quantitated by MTT (3-[4, 5 dimethylthiazol-y-yl]-2, 5-diphenyl tetrazolium bromide) activity measured 4 hours after the addition of MTT labeling reagent to the cell compartment.
RESULTS
At a constant [bilirubin]/[albumin] ratio of 3: 1, the increasing concentration of bilirubin (50microM, 75microM, 100microM, 150microM) resulted in proportionally increased cytotoxicity (18+/-2%, 33+/-1%, 44+/-2%, 66+/-4%, respectively). At a constant bilirubin concentration of 86microM, the increasing [bilirubin]/[albumin] ratio (1: 3, 3: 1) also resulted in proportionally increased cytotoxicity (10+/-1%, 58+/-1%, respectively) and proportionally decreased viability (83+/-1%, 65+/-2%, respectively).
CONCLUSION
Not only the concentration of bilirubin, but also the [bilirubin]/[albumin] ratio may be important for the cerebral cortial cell injury by bilirubin.

Key Words: Bilirubin cytotoxicity; Neuronal cell culture; Hyperbilirubinemia; Kernicterus; Albumin