폴리스티렌 나노섬유상에서의 간세포의 3차원 배양

김영진; 안창현; 오환희; 김영진; 윤관한; 강인규; 권오형; Young-Jin Kim; Chang Hyun Ahn; Hwan Hee Oh; Young-Jin Kim; Kwan Han Yoon; Inn-Kyu Kang; Oh Hyeong Kwon

Polymer(Korea), Vol.32, No.2, 131-137, March, 2008

Export Citation

폴리스티렌 나노섬유상에서의 간세포의 3차원 배양

3-D Rat Hepatocytes' Culture on Polystyrene Nanofibrous Scaffold

김영진, 안창현¹, 오환희², 김영진², 윤관한 ², 강인규 ³, 권오형 ^{2, †}

대구가톨릭대학교 의공학과
¹나노부품실용화센터
²금오공과대학교 고분자공학과
³경북대학교 고분자공학과

Young-Jin Kim, Chang Hyun Ahn¹, Hwan Hee Oh², Young-Jin Kim², Kwan Han Yoon², Inn-Kyu Kang³, and Oh Hyeong Kwon^{2, †}

Department of Biomedical Engineering, Catholic University of Daegu, Daegu 712-702, Korea
¹Nano Practical Application Center, Daegu 704-801, Korea
²Department of Polymer Science and Engineering, Kumoh National Institute of Technology, Gumi 730-701, Korea
³Department of Polymer Science, Kyungpook National University, Deagu 702-701, Korea

E-mail:

초록

전기방사법에 의해 폴리스테렌 나노섬유 부직포를 제조하였고, 간세포의 접착을 유도하기 위해 간세포접착 리간드인 poly[N-p-vinylbenzyl-O-β-D-galactopyranocyl-(1→4)-D-gluconamide](PVLA)를 나노섬유의 표면에 코팅하였다. 얻어진 나노섬유는 평균직경이 500 nm이었고 분광학적인 방법을 통해 PS 나노섬유 표면의 개질 여부를 확인하였다. 장기간 배양시의 간세포 생존율을 MTT법으로 조사한 결과 기존의 세포배양접시에서는 급격하게 세포생존율이 감소함에 비해 PVLA가 코팅된 나노섬유상에서는 생존율이 서서히 감소하였으며 약 3주간에 걸쳐 간세포가 생존율이 유지되었다. 간세포의 대표적인 기능인 알부민분비 및 요소분해능을 측정한 결과 기존의 세포배양접시에서는 급격하게 간세포기능이 저하되었지만, PVLA가 코팅된 나노섬유상에서는 약 3주정도 초기의 간세포기능을 유지하는 것이 밝혀졌다.

We have fabricated polystyrene (PS) nanofibrous scaffold for hepatocyte culture by electrospinning method and subsequently coated with specific ligand of poly[N-p-vinylbenzyl-O-β-D-galactopyranosyl-(1→4)-D-gluconamide](PVLA) to enhance hepatocytes attachment. Rat hepatocytes behavior on the PVLA-coated and non-coated PS nanofibrous matrices have been investigated. Electrospun PS nanofiber structures revealed randomly aligned fibers with average diameter of 500 nm. It is observed that PS nanofibrous matrix could incorporate many cells into the interior of the matrix probably due to the suitable pore size. Cell viabilities cultured on PVLA-coated PS nanofibrous mats were maintained for 3 weeks, while it was decreased rapidly on PVLA-coated PS dishes. High hepatic functions especially for albumin secretion and ammonia removal were maintained at least for 2 weeks on nanofibrous mats but rapidly decreased on flat PS dishes. These results indicate that nanofibrous structure enabled 3-D culture with high level of cell-cell contact results in providing cell-cell communications and subsequent long-term maintenance of specific cell functions.

Keywords:electrospinning;nanofiber;hepatocyte;3-D culture;specific hepatic functions

[References]

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Mikos AG, Sarakinos G, Leite SM, Vacanti JP, Langer R, Biomaterials, 14, 323 (1993)
Mikos AG, Thorsen AJ, Czerwonka LA, Bao Y, Langer R, Winslow DN, Vacanti JP, Polymer, 35(5), 1068 (1994)
Cho SH, Oh SH, Lee JH, J. Biomater. Sci.-Polym. Ed., 16, 933 (2005)
Ko YK, Kim SH, Jeong JS, Ha HJ, Yoon SJ, Rhee JM, Kim MS, Lee HB, Khang G, Polym.(Korea), 31(1), 14 (2007)
Langer R, Vacanti JP, Science, 260, 920 (1993)
Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S, Compos. Sci. Technol., 63, 2223 (2003)
Fong H, Chun I, Reneker DH, Polymer, 40(16), 4585 (1999)
Zong XH, Kim K, Fang DF, Ran SF, Hsiao BS, Chu B, Polymer, 43(16), 4403 (2002)
Lee IS, Kwon OH, Meng W, Kang IK, Ito Y, Macromol. Res., 12(4), 374 (2004)
Bhatia SN, Yarmush ML, Toner M, J. Biomed. Mater. Res., 34, 189 (1997)
Yamato M, Kwon OH, Hirose M, Kikuchi A, Okano T, J. Biomed. Mater. Res., 55, 137 (2001)
Hoshiba T, Wakejima M, Cho CS, Shiota G, Akaike T, Biomed J, Mater. Res. A, in press
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Geng X, Kwon OH, Jang J, Biomaterials, 26, 5427 (2005)
Ito Y, Hasuda H, Kamitakahara M, Otsuki C, Tanihara M, Kang IK, Kwon OH, J. Bio. Bioeng., 100, 43 (2005)
Kwon OH, Lee IS, Ko YG, Meng W, Jung KH, Kang IK, Ito Y, Biomedial Materials, 2, S25 (2007)
Jin HJ, Chen J, Karageorgiou V, Altman GH, Kaplan DL, Biomaterials, 25, 1039 (2004)
Meng W, Kim SY, Yuan J, Kim JC, Kwon OH, Kawazoe N, Chen G, Ito Y, Kang IK, J. Biomater. Sci. Polym. Ed., 18, 81 (2007)

[Related Articles]

[1] Vacanti JP, Craniofacial Growth Series, 34, 171 (1998)

[2] Fuller B, J. Hepatol., 7, 368 (1988)

[3] Rivas P, Fabrega A, Transplant. Proc., 24, 1833 (1992)

[4] Mikos AG, Sarakinos G, Leite SM, Vacanti JP, Langer R, Biomaterials, 14, 323 (1993)

[5] Mikos AG, Thorsen AJ, Czerwonka LA, Bao Y, Langer R, Winslow DN, Vacanti JP, Polymer, 35(5), 1068 (1994)

[6] Cho SH, Oh SH, Lee JH, J. Biomater. Sci.-Polym. Ed., 16, 933 (2005)

[7] Ko YK, Kim SH, Jeong JS, Ha HJ, Yoon SJ, Rhee JM, Kim MS, Lee HB, Khang G, Polym.(Korea), 31(1), 14 (2007)

[8] Langer R, Vacanti JP, Science, 260, 920 (1993)

[9] Huang ZM, Zhang YZ, Kotaki M, Ramakrishna S, Compos. Sci. Technol., 63, 2223 (2003)

[10] Fong H, Chun I, Reneker DH, Polymer, 40(16), 4585 (1999)

[11] Zong XH, Kim K, Fang DF, Ran SF, Hsiao BS, Chu B, Polymer, 43(16), 4403 (2002)

[12] Lee IS, Kwon OH, Meng W, Kang IK, Ito Y, Macromol. Res., 12(4), 374 (2004)

[13] Bhatia SN, Yarmush ML, Toner M, J. Biomed. Mater. Res., 34, 189 (1997)

[14] Yamato M, Kwon OH, Hirose M, Kikuchi A, Okano T, J. Biomed. Mater. Res., 55, 137 (2001)

[15] Hoshiba T, Wakejima M, Cho CS, Shiota G, Akaike T, Biomed J, Mater. Res. A, in press

[16] Akaike T, Tobe S, Kobayashi A, Goto M, Kobayashi K, Gastroenterol. Jpn., 28, 45 (1993)

[17] Geng X, Kwon OH, Jang J, Biomaterials, 26, 5427 (2005)

[18] Ito Y, Hasuda H, Kamitakahara M, Otsuki C, Tanihara M, Kang IK, Kwon OH, J. Bio. Bioeng., 100, 43 (2005)

[19] Kwon OH, Lee IS, Ko YG, Meng W, Jung KH, Kang IK, Ito Y, Biomedial Materials, 2, S25 (2007)

[20] Jin HJ, Chen J, Karageorgiou V, Altman GH, Kaplan DL, Biomaterials, 25, 1039 (2004)

[21] Meng W, Kim SY, Yuan J, Kim JC, Kwon OH, Kawazoe N, Chen G, Ito Y, Kang IK, J. Biomater. Sci. Polym. Ed., 18, 81 (2007)