High-Throughput Single-Cell Derived Sphere Formation for Cancer Stem-Like Cell Identification and Analysis.
Considerable evidence suggests that many malignancies are driven by a cellular compartment that displays stem cell properties. Cancer stem-like cells (CSCs) can be identified by expression of cell surface markers or enzymatic activity, but these methods are limited by phenotypic heterogeneity and plasticity of CSCs. An alternative phenotypic methodology based on in-vitro sphere formation has been developed, but it is typically labor-intensive and low-throughput. In this work, we present a 1,024-microchamber microfluidic platform for single-cell derived sphere formation. Utilizing a hydrodynamic capturing scheme, more than 70% of the microchambers capture only one cell, allowing for monitoring of sphere formation from heterogeneous cancer cell populations for identification of CSCs. Single-cell derived spheres can be retrieved and dissociated for single-cell analysis using a custom 96-gene panel to probe heterogeneity within the clonal CSC spheres. This microfluidic platform provides reliable and high-throughput sphere formation for CSC identification and downstream clonal analysis.
Department of Electrical Engineering and Computer Science, University of Michigan, 1301 Beal Avenue, Ann Arbor, MI 48109-2122, USA.|University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Drive, Ann Arbor, MI 48109-5940, USA.
Yu-Chih
YC
Chen
Department of Biomedical Engineering, University of Michigan, 2200 Bonisteel Blvd, Ann Arbor, MI 48109-2099, USA.
Patrick N
PN
Ingram
University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Drive, Ann Arbor, MI 48109-5940, USA.
Shamileh
S
Fouladdel
University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Drive, Ann Arbor, MI 48109-5940, USA.
Sean P
SP
McDermott
University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Drive, Ann Arbor, MI 48109-5940, USA.
Ebrahim
E
Azizi
University of Michigan Comprehensive Cancer Center, 1500 East Medical Center Drive, Ann Arbor, MI 48109-5940, USA.
Max S
MS
Wicha
Department of Electrical Engineering and Computer Science, University of Michigan, 1301 Beal Avenue, Ann Arbor, MI 48109-2122, USA.|Department of Biomedical Engineering, University of Michigan, 2200 Bonisteel Blvd, Ann Arbor, MI 48109-2099, USA.
Euisik
E
Yoon
D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D061986:MCF-7 Cells; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D011102:Polyhydroxyethyl Methacrylate / Q000737:chemistry; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods; D018874:Spheroids, Cellular / Q000166:cytology* / Q000378:metabolism; D014183:Transplantation, Heterologous
D014408:Biomarkers, Tumor; D020100:Hydrogels; D051880:Receptors, Notch; D011102:Polyhydroxyethyl Methacrylate