1. A Perfusion-based Micro 3-D Cell Culture Platform NTHU MSCL Adviser: C. H. Liu C. He Yeh

2. A Perfusion-based Micro 3-D Cell Culture Platform Outline  Culture environment at large scale  Culture environment at micro scale  Principle  Fabrication  Conclusion & Features  Further work

3. General Culture Equipment 細胞培養設備 (1) 高溫高壓滅菌爐 (2) 無菌室 (3) 無菌操作台 (4) 二氧化碳培養箱 (5) 細胞離心機 (6) 倒立顯微鏡 (7) 恆溫水浴槽 (8) 冷凍冷藏冰箱 (9) 細胞計數器及血球計數盤 (10) 本生燈 (11) 液氮細胞儲存桶 (12) 酸鹼測定儀 (13) 真空幫浦 (14) 瓦斯桶 (15) 二氧化碳桶 Lab On a Chip

4. On Chip Advantages Small dimension Saving sample Rapid assay process (micro dimension, avoid medium or reagent diffusion time gradient or concentration different ) Cheap, disposable

5. On Chip Culture Requirement Temperature : 37 ℃ pH : 7.2 ~ 7.4 ( 95% air 5%CO 2 or HEPES) Buffered saline (PBS, PBSA, …etc) Medium : (Serum, glucose, amino acids, vitamin, antibiotics ) 3D culture (In vivo Culture)

6. A Perfusion-based Micro 3-D Cell Culture Platform

7. Working Principle :

8. A Perfusion-based Micro 3-D Cell Culture Platform Fabrication :

9. Conclusion Feature 1: 3D well-defined culture environment  mimic an in vivo environment  distinct cellular behaviors (cell-cell and cell-extracellular interactions )  provide more meaningful data

10. Conclusion Feature 2 : Short Loading cell channel Feature 3 : High throughput (8 chambers)

11. Further work Bioassay (inject reagent instead of culture medium in vivo bioassay in vivo bioassay) Biochemical experiment Tissue engineering Drug screening μTAS

12. Reference “A PERFUSION-BASEDMICRO 3-D CELL CULTURE PLATFORM” S. B. Huang1, M. H. Wu2, Z. F. Cui3, Z. Cui2, J. L. Lin4 and G. B. Lee1* “MICROFLUIDIC DEVICES FOR THE ASSEMBLY AND CULTURE OF THREE-DIMENSIONAL MULTI- CELLULAR CONSTRUCTS WITH DIFFUSION-LIMITED MICROENVIRONMENTS” M. Pinelis1, R. W. Kasinskas2, R. T. Borno1, J. H. Park1, E. Chu3, N. S. Forbes2 and M. M. Maharbiz1 "Differentiation of human Embryonic Stem Cells into Hepatocytes in 2D and 3D culture system in vitro" The International Journal of Developmental Biology, Vol. 50, pp. 645-652, 2006 "The multilayered postconfluent cell culture as a model for drug screening" Critical Reviews in Oncology/Hematol, Vol. 36, pp. 147-157, 2000. J. M. Pardon; C.L. van der Wilt; K. Smid; E. Smitskamp-Wilms; H.H. Baukus; P. E. Piazo; G. "Development of PDMS Microbioreactor with Well-defined and Homogenous Culture Environment for Chondrocyte 3-D Culture“ M. H. Wu, J. P. G. Urban, Z. Cui and Z. F. Cui, Biomedical Microdevices, Vol. 8, pp. 331-340, 2006.