結言

本章では、培養される各臓器の機能の再現、各細胞の培養環境の相違、システムの 大型化、流路への代謝物質の吸着と吸収の解決を目的とし、細胞数を考慮した細胞培 養チャンバの設計、モジュラ機構、重力送液、無吸収材料流路を特色とする

Body-on-a-Chip用プラットフォームの設計、作製と評価を行った。

培養される各臓器の機能再現の観点から、各臓器の細胞数を考慮した流体システム 設計を行った。また、各細胞の培養環境の相違を解決するために、初期培養環境を個 別に設定できるモジュラ機構を組み込んだ。また、システムの大型化を回避するため に、重力送液を提案した。さらに、流路への代謝物質の吸着と吸収を抑制するために、

無吸収材料のみを使用したマイクロ流路を作製した。以上の機能を組み込んだ提案プ ラットフォームを用いることにより、それぞれ異なる環境で培養した細胞を、共培養 環境に格納後、24 h共培養した細胞からの蛍光を取得した。培養後の観察においても 細胞の接着が見られたことから、提案するロッカープラットフォームは、各培養チャ ンバに十分な培地量を送液できたといえる。今後は、アルブミンや尿素などの細胞の 代謝物質濃度と、実際に播種された細胞の数を比較し、正常な代謝活動の評価と、複 数種類の細胞の共培養を行うことで、プラットフォームのBody-on-a-Chip向けの有効 性を検証することができる。その際に、本論文の3章で提案した立体的に細胞構造を 再現できるスカフォールドを格納し、同じく代謝物質の比較を行うことで、より正確 な細胞代謝機能のアッセイに有効であると考えられる。

156

参考文献

[1] U. Marx, T.B. Andersson, A. Bahinski, M. Beilmann, S. Beken, F.R. Cassee, M. Cirit, M. Daneshian, S. Fitzpatrick, O. Frey, C. Gaertner, C. Giese, L. Griffith, T. Hartung, M.B. Heringa, J. Hoeng, W.H. Jong, H. Kojima, J. Kuehnl, A. Luch, I. Maschmeyer, D.

Sakharov, A.J. Sips, T. Steger-Hartmann, D.A. Tagle, A. Tonevitsky, T. Tralau, S. Tsyb, A. Stolpe, R. Vandebriel, P. Vulto, J. Wang, J. Wiest, M. Rodenburg, and A. Roth,

“Biology-inspired microphysiological system approaches to solve the prediction dilemma of substance testing”, Alternatives to Animal Experimentation, Vol. 33, pp.

272-321, 2016.

[2] E. Dehne, T. Hasenberg, and U. Marx, “The ascendance of microphysiological systems to solve the drug testing dilemma”, Future Science, Vol. 3, pp. 2056-5623, 2017.

[3] M.B. Esch, A.S. Smith, J. Prot, C. Oleaga, J.J. Hickman, and M.L. Shuler, “How multi-organ microdevices can help foster drug development”, Advanced Drug Delivery, Vol.

69, pp. 158-169, 2014.

[4] Y.I. Wang, C. Oleaga, C.J. Long, M.B. Esch, C.W. McAleer, P.G. Miller, J.J. Hickman, and M.L. Shuler, “Self-contained, low-cost body-on-a-chip systems for drug development”, Experimental Biology and Medicine, Vol. 242, pp. 1701-1713, 2017.

[5] U. Marx, H. Walles, S. Hoffmann, G. Lindner, R. Horland, F. Sonntag, U. Klotzbach, D.

Sakharov, A. Tonevitsky, and R. Lauster, “‘Human-on-a-chip’ developments: a translational cuttingedge alternative to systemic safety assessment and efficiency evaluation of substances in laboratory animals and man?”, Alternatives to Laboratory Animals, Vol. 40, pp. 235-257, 2012.

[6] N.S. Bhise, J. Ribas, V. Manoharan, Y.S. Zhang, A. Polini, S. Massa, M.R. Dokmeci, and A. Khademhosseini, “Organ-on-a-chip platforms for studying drug delivery systems”, Journal of Controlled Release, Vol. 190, pp. 82-93, 2014.

[7] C. Moraes, G. Mehta, S.C. Lesher-Perez, and S. Tkakayama, “Organs-on-a-chip: a focus on compartmentalized microdevices”, Annals of Biomedical Engineering, Vol. 40, pp.

1211-1227, 2012.

[8] C.D. Edington, W.L.K. Chen, E. Geishecker, T. Kassis, L.R. Soenksen, B.M. Bhushan, D. Freake, J. Kirschner, C. Maass, N. Tsamandouras, J. Valdez, C.D. Cook, T. Parent, S. Snyder, J. Yu, E. Suter, M. Shockley, J. Velazquez, J.J. Velazquez, L. Stockdale, J.P.

157

Papps, I. Lee, N. Vann, M. Gamboa, M.E. Labarge, Z. Zhong, X. Wang, L.A. Boyer, D.A. Lauffenburger, R.L. Carrier, C. Communal, S.R. Tannenbaum, C.L. Stokes, D.J.

Hughes, G. Rohatgi, D.L. Trumper, M. Cirit, and L.G. Griffith, “Interconnected microphysiological systems for quantitative biology and pharmacology studies”, Scientific Reports, Vol. 8, pp. 1-18, 2018.

[9] I. Wagner, E. Materne, S. Brincker, U. Su, C. Fradrich, M. Busek, F. Sonntag, D.A.

Sakharov, E.V. Trushkin, A.G. Tonevitsky, R. Laustera, and U. Marx, “A dynamic multi-organ-chip for long-term cultivation and substance testing proven by 3D human liver and skin tissue co-culture”, Lab on a Chip, Vol. 13, pp. 3538-3547, 2013.

[10] P.G. Miller and M.L. Shuler, “Design and demonstration of a pumpless 14 compartment microphysiological system”, Biotechnology and Bioengineering, Vol. 113, pp. 2213-2227, 2016.

[11] S. Shay, “Organs-on-a-chip: a future of rational drug-design”, Journal of Biosciences and Medicines, Vol. 5, pp. 22-28, 2017.

[12] E. Bianconi, A. Piovesan, F. Facchin, A. Beraudi, R. Casadei, F. Frabetti, L. Vitale, M.C.

Pelleri, S. Tassani, F. Piva, S. Amodio, P. Strippoli, and S. Canaider, “An estimation of the number of cells in the human body”, Annals of Human Biology, Vol. 40, pp. 463-471, 2013.

[13] Y. Higashi, K. Noma, M. Yoshizumi, and Y. Kihara, “Endothelial functionand in cardiovascular oxidative stress diseases”, Citrculation Journal, Vol. 73, pp. 411-418, 2009.

[14] M.W. Stefness, D. Schmidt, R. Mccrery, and J.M. Basgen, “Glomerular cell number in normal subjects and in type 1 diabetic patients”, Kidney International, Vol. 59, pp. 2104-2113, 2001.

[15] P.S. Price, R.B. Conolly, C.F. Chaisson, E.A. Gross, J.S. Young, E.T. Mathis, and D.R.

Tedder, “Modeling interindividual variation in physiological factors used in PBPK models of humans”, Critical Reviews in Toxicology, Vol. 33, pp. 469-503, 2003.

[16] H.F. Helander and L. Fandriks, “Surface area of the digestive tract-revisited”, Scandinavian Journal of Gastroenterology, Vol. 49, pp. 681-689, 2014.

[17] B. Davies and T. Morris, “Physiological parameters in laboratory animals and humans”, Pharmaceutical Research, Vol. 10, pp. 1093-1095, 1993.

158

[18] N. Tsuchiya, T. Yamashiro, and S. Murayama, “Decrease of pulmonary blood flow detected by phase contrast MRI is correlated with a decrease in lung volume and increase of lung fibrosis area determined by computed tomography in interstitial lung disease”, European Journal of Radiology, Vol. 85, pp. 1581-1585, 2016.

[19] W.G. Schenk, J.C. Mcdonald, K. Mcdonald, and T. Drapanas, “Direct measurement of hepatic blood flow in surgical patients: with related observations on hepatic flow dynamics in experimental animals”, Annals of Surgery, Vol. 156, pp. 463-469, 1962.

[20] J.H. Sung, M.B. Esch, J. Prot, C.J. Long, A. Smith, J. Hickman, and M.L. Shuler,

“Microfabricated mammalian organ systems and their integration into models of whole animals and humans”, Lab on a Chip, Vol. 13, pp. 1201-1212, 2013.

[21] K.W. Oh, K. Lee, B. Ahna, and E. P. Furlani, ”Design of pressure-driven microfluidic networks using electric circuit analogy”, Lab on a Chip, Vol. 12, pp. 515-545, 2012.

[22] J. Yeom, Y. Wu, and M.A. Shannon, “Maximum achievable aspect ratio in deep reactive ion etching of silicondue to aspect ratio dependent transport and the microloading effect”, American Vacuum Society, Vol. 23, pp. 2319-2329, 2005.

[23] R. Baudoin, L. Griscom, J.M. Prot, C. Legallais, and E. Leclerc, “Behavior of HepG2/C3A cell cultures in a microfluidic bioreactor”, Biochemical Engineering Journal, Vol. 53, pp. 172-181, 2011.

[24] J.H. Yeon and J. Park, “Cytotoxicity test based on electrochemical impedance measurement of HepG2 cultured in microfabricated cell chip”, Analytical Biochemistry, Vol. 341, pp. 308-315, 2005.

159