High-throughput stem cell-based phenotypic screening through microniches

• Published in: Biomaterials science Vol. 7; no. 8; pp. 3471 - 3479
• Main Authors: Kolb, Laura, Allazetta, Simone, Karlsson, Maria, Girgin, Mehmet, Weber, Wilfried, Lutolf, Matthias P
• Format: Journal Article
• Language: English
• Published: England Royal Society of Chemistry 23.07.2019
• Subjects: Animals; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biotechnology; Cell Line; Fluorescence; Gene sequencing; Glycoproteins - genetics; Glycoproteins - metabolism; Growth factors; HEK293 Cells; High-Throughput Screening Assays - instrumentation; High-Throughput Screening Assays - methods; Humans; Immunoglobulin Fc Fragments - genetics; Immunoglobulin Fc Fragments - metabolism; Intercellular Signaling Peptides and Proteins - genetics; Intercellular Signaling Peptides and Proteins - metabolism; Interleukin-4 - genetics; Interleukin-4 - metabolism; Interleukins; Lab-On-A-Chip Devices; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Microgels; Polymerization; Protein Engineering - methods; Proteins; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Screening; Sequence Analysis, RNA; STAT6 Transcription Factor - genetics; STAT6 Transcription Factor - metabolism; Stem cells; Stem Cells - cytology; Stem Cells - physiology; Tissue engineering; Workflow
• ISSN: 2047-4830; 2047-4849
• DOI: 10.1039/c8bm01180j

As the field of tissue engineering develops, methods for screening combinations of signals for their effects on stem cell behavior are needed. We introduce a microgel-based screening platform for testing combinations of in situ -generated proteins on stem cell fate in ultrahigh-throughput. Compartmentalizing individual sets of growth factors was addressed by encapsulating aggregates of stable recombinant cell lines secreting individual glycoproteins into microgels through an on-chip polymerization. When these 'microniches' are cultured with a cell type of interest, fluorescence reporters indicate positive niches that perform the desired function, and the underlying producer cell lines of these selected microniches are analyzed by barcoded RNA sequencing. The microniche-based screening work-flow was validated via a model system based on engineered mammalian cells expressing yellow fluorescent protein (YFP) upon anti-inflammatory cytokine interleukin 4 (IL4)-based activation. Methods for screening combinations of signals for their effects on stem cell behavior are needed in the field of tissue engineering. We introduce a microgel-based screening platform for testing combinations of proteins on stem cell fate.