RNA-Seq Blog Transcriptome Research & Industry News
Tissues are complex mixtures of different cell subtypes, and this diversity is increasingly characterized using high-throughput single cell analysis methods. However, these efforts are hindered, as tissues must first be dissociated into single cell suspensions using methods that are often inefficient, labor-intensive, highly variable, and potentially biased towards certain cell subtypes.
University of California, Irvine researchers present a microfluidic platform consisting of three tissue processing technologies that combine tissue digestion, disaggregation, and filtration. The platform is evaluated using a diverse array of tissues. For kidney and mammary tumor, microfluidic processing produces 2.5-fold more single cells. Single cell RNA sequencing further reveals that endothelial cells, fibroblasts, and basal epithelium are enriched without affecting stress response. For liver and heart, processing time is dramatically reduced. The researchers also demonstrate that recovery of cells from the system at periodic intervals during processing increases hepatocyte and cardiomyocyte numbers, as well as increases reproducibility from batch-to-batch for all tissues.
Microfluidic tissue processing platform
a Schematic of the minced tissue digestion device. Design includes six total layers, including two fluidic layers (green), 2 via layers (red), and the top and bottom end caps (gray). Tissue is loaded through the luer port and into the tissue chamber. b Schematic of the tissue chamber. Fluidic channels direct hydrodynamic shear forces and proteolytic enzymes, while also retaining minced tissue pieces in the chamber. c Schematic of the integrated dissociation/filter device. Tissue fragments and cell aggregates from the digestion device will be further broken down by hydrodynamic shear forces and nylon mesh filters. d Pictures of the fabricated minced digestion device. e Picture of the fabricated dissociation/filter device.
