3D-Printing Microfludic Device for Breast Cancer Cell Culture and Pharmacological Research
LU Yong-zhuang1, REN Yi-na1, GONG Ming-hua2, YANG Bing-xian1, CAO Ming3, ZHANG Lin1*
1.College of Biomedical Engineering and Instrument Science, Zhejiang University, Hangzhou 310027, China; 2.School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China; 3. School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, China
Abstract��OBJECTIVE To develop a microfluidic mammalian cell culture device with human breast cancer cells co-cultured with extracellular matrix (ECM), reconstitute the 3-dimension human body microenvironment and preliminarily conduct drug screening on the breast cancer chip. METHODS 3D Printing technology was utilized to build a two-layer microchip with cell culture reservoirs. The breast cancer cells (MCF7) were cultured in the matrix gel structure which mimiced the 3-dimension human body microenvironment. The cell toxicity of paclitaxel (PTX) on MCF7 cells was preliminarily observed using this microfluidic chip. With the use of AO/EB immunofluorescent staining and laser confocal microscopy, the cell death ratio induced by PTX was determined and compared with that determined by 2-dimension drug screening methods. RESULTS MCF7 cells cultured on the chip successfully formed a 3D cavity structure in the matrix after 6-day dynamic incubation. The flow rate was 10 ��L��h-1. After 24 h dynamic culture, the culture medium was changed to culture solution containing 0.02 ��mol��mL-1 PTX, and the incubation was continued for 24 h. Obvious cell death was detected under laser confocal microscopy. CONCLUSION The microfluidic chip developed in this study can successfully culture breast cancer cells in 3-dimension structure and perform drug screening, which lays a foundation for actualization of ��human-on-a-chip��.
¬Ӿׯ, ������, ������, �����, ����, ����. ����3D��ӡ����������оƬ�������ҩЧɸѡ[J]. �й�ҩѧ��־, 2015, 50(24): 2124-2129.
LU Yong-zhuang, REN Yi-na, GONG Ming-hua, YANG Bing-xian, CAO Ming, ZHANG Lin. 3D-Printing Microfludic Device for Breast Cancer Cell Culture and Pharmacological Research. Chinese Pharmaceutical Journal, 2015, 50(24): 2124-2129.
ESCH M B, KING T L, SHULER M L. The role of body-on-a-chip devices in drug and toxicity studies[J]. Annu Rev Biomed Eng, 2011, 13: 55-72.
[2]
SWEENEY L M, SHULER M L, BABISH J G, et al. A cell culture analogue of rodent physiology: Application to naphthalene toxicity[J]. Toxicol In Vitro, 1995, 9(3):307-316.
[3]
LEE M Y, KUMAR R A, SUKUMARAN S M, et al. Three-dimensional cellular microarray for high-throughput toxicology assays[J]. Proc Natl Acad Sci USA, 2008, 105(1):59-63.
[4]
PAULL K D, SHOEMAKER R H, HODES L, et al. Display and analysis of patterns of differential activity of drugs against human tumor cell lines: Development of mean graph and COMPARE algorithm[J]. J Natl Cancer Inst, 1989, 81(14): 1088-1092.
[5]
JOHNSON J I, DECKER S, ZAHAREVITZ D, et al. Relationships between drug activity in NCI preclinical in vitro and in vivo models and early clinical trials[J]. Br J Cancer, 2001, 84(10): 1424.
[6]
WHIESIDES G M. The origins and the future of microfluidics[J]. Nature, 2006, 442(7101): 368-373.
[7]
ABACI H E, GLEDHILL K, GUO Z, et al. Pumpless microfluidic platform for drug testing on human skin equivalents[J]. Lab Chip, 2015,15(3):882-888.
[8]
AU S H, CHAMBERLAIN M D, MAHESH S, et al. Hepatic organoids for microfluidic drug screening[J]. Lab Chip, 2014, 14(17): 3290-3299.
[9]
MINSEOK S K, JU H Y, JE-KYUN P.A microfluidic platform for 3-dimensional cell culture and cell-based assays[J]. Biomed Microdevices, 2007, 9(1):25-34.
[10]
JANG K J, SUH K Y. A multi-layer microfluidic device for efficient culture and analysis of renal tubular cells[J]. Lab Chip, 2010, 10(1): 36-42.
[11]
HUH D, MATTEWS B D, MAMMOTO A, et al. Reconstituting organ-level lung functions on a chip[J]. Science, 2010, 328(5986): 1662-1668.
[12]
KIM H J, HUH D, HAMILTON G, et al. Human gut-on-a-chip inhabited by microbial flora that experiences intestinal peristalsis-like motions and flow[J]. Lab Chip, 2012, 12(12): 2165-2174.
[13]
ZHANG Q, ZHOU X M, YAN W, et al. Three dimensional cultures of liver cancer cell on a poly(dimethyl siloxane)-paper hybrid microfluidic chip[J]. Chin J Anal Chem(������ѧ), 2012, 40(7): 996-1001.
[14]
ZHENG Y H, WU J Z, SHAO J B, et al. Microfluidic cell culture array chip for drug screening assays[J]. Chin J Biotechnol(���﹤��ѧ��), 2009, 25(5): 779-785.
[15]
SONG Z, CAI S X, CHEN S J, et al. A novel cancer invasion model based on a microfluidic device in a 3-dimensional matrix (in Chinese)[J]. Chin Sci Bull (��ѧͨ��), 2012, 57(23): 2184-2191.
[16]
BHATIA S N, BALIS U J, YARMUSH M L, et al. Effect of cell-cell interactions in preservation of cellular phenotype: Cocultivation of hepatocytes and nonparenchymal cells[J]. FASEB J, 1999, 13(14):1883-1900.
[17]
KUMAR N M,GILULA N B. The gap junction communication channel[J]. Cell, 1996, 84(3):381-388.
[18]
PAWSON T. Protein modules and signalling networks [J]. Nature,1995,373(6515):573-580.
[19]
WISEMAN B S, WERB Z. Effects on mammary gland development and breast cancer[J].Science,2002,296(5570):1046-1049.
[20]
BISSELL M J, RADISKY D. Putting tumors in context[J]. Nat Rev Cancer, 2001,1(1):46-54.
[21]
SUNG K E. Control of 3-dimensional collagen matrix polymerization for reproducible human mammary fibroblast cell culture in microfluidic devices[J]. Biomaterials, 2009, 30(27): 4833-4841.
[22]
BAUDOIN R, GRISCOM L, MONGE M, et al. Development of a renal microchip for in vitro distal tubule models[J]. Biotechnol Prog, 2007,23(5): 1245-1253.
[23]
BLAGOVIC K, KIM L Y, VOLDMAN J. Microfluidic perfusion for regulating diffusible signaling in stem cells[J]. PLoS One, 2011, 6(8):e22892.
[24]
HAYES A W. Principles and Methods of Toxicology[M]. 4th ed; Philadelphia:Taylor and Francis, 2001.
[25]
GUILLOUZO. A. Liver cell models in in vitro toxicology[J]. EnViron Health Perspect,1998, 106 (2):511-532.
[26]
MCCORMICK R M, NELSON R J, ALONSO-AMIGO M G, et al. Microchannel electrophoretic separations of DNA in injection-molded plastic substrates[J]. Anal Chem, 1997, 69(14): 2626-2630.
[27]
HUH D, HAMILTON G A, INGBER D E. From 3D cell culture to organs-on-chips[J]. Trends Cell Biol, 2011, 21(12): 745-754.
[28]
SCHUBERT C, VAN L M C, DONOSO L A. Innovations in 3D printing: A 3D overview from optics to organs[J]. Br J Ophthalmol, 2014,98(2):159-161.
[29]
KLEIN G T, LU Y, WANG M Y. 3D Printing and neurosurgery��ready for prime time?[J]. World Neurosurg, 2013,80(3-4):233-235.
[30]
LEE C, VENTOLA M S. Medical applications for 3D printing: Current and projected uses[J]. P T,2014,39(10): 704-711.
[31]
MIRONOV V, KASYANOV V, DRAKE C, et al. Organ printing: Promises and challenges[J]. Regen Med, 2008,3 (1) :93-103.
[32]
GAO Q, HE Y, FU J Z, et al. Coaxial nozzle-assisted 3D bioprinting with built-in microchannels for nutrients delivery[J]. Biomaterials, 2015,61:203-215.
[33]
SEIDMAN A D, TIERSTEN A, HUDIS C, et al. Phase II trial of paclitaxel by 3-hour infusion as initial and salvage chemotherapy for metastatic breast cancer[J]. J Clin Oncol, 1995, 13 (10):2575-2581.
2015, Vol. 50 
