Universiti Malaysia Sabah 1 article published in JoVE Bioengineering Fabrication of a Multiplexed Artificial Cellular MicroEnvironment Array Yasumasa Mashimo1,2, Momoko Yoshioka1, Yumie Tokunaga1, Christopher Fockenberg1, Shiho Terada1, Yoshie Koyama1, Teiko Shibata-Seki2, Koki Yoshimoto1, Risako Sakai1, Hayase Hakariya1, Li Liu1, Toshihiro Akaike3, Eiry Kobatake2, Siew-Eng How4, Motonari Uesugi1,5, Yong Chen1,6, Ken-ichiro Kamei1 1Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, 2Department of Life Science and Technology, School of Life Science and Technology, Tokyo Institute of Technology, 3Biomaterials Center for Regenerative Medical Engineering, Foundation for Advancement of International Science, 4Faculty of Science and Natural Resources, Universiti Malaysia Sabah, 5Institute for Chemical Research, Kyoto University, 6Ecole Normale Supérieure This article describes the detailed methodology to prepare a Multiplexed Artificial Cellular MicroEnvironment (MACME) array for high-throughput manipulation of physical and chemical cues mimicking in vivo cellular microenvironments and to identify the optimal cellular environment for human pluripotent stem cells (hPSCs) with single-cell profiling.