title: High-throughput superhydrophobic microwell arrays for investigating multifactorial stem cell niches.
abstract: Understanding the complex regulatory network that determines stem cell fates requires a high-throughput platform that can generate a large number of precisely controlled microenvironments representing multiple factors for stem cell culture and analysis. Here, we developed a superhydrophobic microwell array chip on which the culture conditions in each microwell can be spontaneously isolated by a grafted layer of superhydrophobic polymers. Simple steps for medium exchange were developed to facilitate the on-chip culture of both adherent and non-adherent cells for up to six days without compromising cell viability and functionality. The culture conditions in each microwell were facilely manipulated using a robotic spotter. Stem cell niches combining soluble factors, extracellular matrices and microtopographic cues were generated on a single 512-well SMARchip and their combinatorial effects on the fate of mouse Oct4-EGFP iPSCs were systematically probed. We observed significant differences in iPSC pluripotency and proliferation between adherent flat and suspended spherical cultures on our platform, which might provide insights into improvement of stem cell technologies.
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Jianxiong
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Shengtai
authors.initials: S
authors.lastname: Bian
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China. xfchen@buaa.edu.cn.
authors.forename: Zhongyao
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Yawei
authors.initials: Y
authors.lastname: Hu
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China. xfchen@buaa.edu.cn.
authors.forename: Ruowen
authors.initials: R
authors.lastname: Hu
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China. xfchen@buaa.edu.cn.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Yichun
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: National Engineering Research Center for Beijing Biochip Technology, Beijing, 102206, China.
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Zhang
authors.affiliation: National Engineering Research Center for Beijing Biochip Technology, Beijing, 102206, China.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: School of Biological Science and Medical Engineering, Beihang University, Haidian District, Beijing, 100191, China. xfchen@buaa.edu.cn.
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Haidian District, Beijing, 100084, China. pliu@tsinghua.edu.cn.
authors.forename: Peng
authors.initials: P
authors.lastname: Liu
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008647:Mesocricetus; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis; D012371:Robotics; D055153:Stem Cell Niche; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins