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. 2020 May 7;8:327. doi: 10.3389/fcell.2020.00327

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Copyright © 2020 Cho, Chiang, Hsieh, Yang, Hsu, Yeh, Huang and Huang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The procedure of forming the hanging drops using the PANDA system. Step 1: Add the cell suspension on the well plate. Step 2: Pour the excess cell suspension into a centrifuge tube. Step 3: Use a scraper to remove the residue. Step 4: Fill the reservoir with PBS through the loading hole. Step 5: Seal the loading hole. Step 6: Connect with the syringe pump to initiate the withdrawal of air and wait until all hanging drops are formed. Step 7: Seal the wells using the sealing sheet. The PANDA system is now ready for cell culture. Step 8: After cell culture, disassemble the chip and collect the hanging drops using a pipette for further experiments. All scale bar represent 2 cm.