while glomerular cell types were enriched to 15.6% (Additional file1: Figure S2e). The fractions of control and DN cells in each cell type were similar except for podocytes, which included more control (approximately 77%, 807 cells) than DN cells (approximately 23%, 235 cells) (Additional ...
The tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC) is a complex ecosystem that drives tumor progression; however, in-depth single cell characterization of the PDAC TME and its role in response to therapy is lacking. Here, we perf
We isolated CD45-negative cells from the single-cell suspension obtained from fresh pancreatic cancer tumor tissues using magnetic-activated cell sorting (MACS; Miltenyi Biotec). After thawing the cryopreserved single-cell suspensions, we depleted cell debris, dead cells, and dying cells using the De...
An aliquot of the singe cell suspension was stained with the LIVE/DEAD™ Viability/Cytotoxicity Kit for mammalian cells (Invitrogen) to ensure viability was greater than 85%. Samples were processed within 3 h from surgical removal to loading on the Chromium (10X Genomics) instrument. We ...
A key aspect of nearly all single-cell sequencing experiments is dissociation of intact tissues into single-cell suspensions. While many protocols have been optimized for optimal cell yield, they have often overlooked the effects that dissociation can ha
The tissues were processed and dissociated into single-cell suspension and sequenced using the Chromium single-cell RNA-Seq platform (10x Genomics). After stringent quality control and normalization analysis, we obtained high-quality transcriptomic profile data from a total of 8000 cells from the 10 ...
In order to study tumor infiltrating B cells (TIBs), the tumor first has to be dissociated into a single cell suspension. For this, the gentleMACS™ Dissociator in combination with the Tumor Dissociation Kit, human ensures rapid and standardized dissociation of tissue samples into single-cell s...
Tissue was digested using dispase, collagenase I, and DNase using the gentle MACS 63 dissociator (Miltenyi) for 30 minutes at 37°C. The cell suspension was passed over 70uM and 40uM filters to generate a single cell suspension. Magnetic bead sorting using MACS LS columns (Miltenyi) and the...
Precancerous lesions of the oral mucosa, especially those accompanied by moderate to severe dysplasia, contribute to the initiation of oral squamous cell carcinoma (OSCC). However, the cellular compositions and spatial organization of the precancerous st
How cell-to-cell copy number alterations that underpin genomic instability1 in human cancers drive genomic and phenotypic variation, and consequently the evolution of cancer2, remains understudied. Here, by applying scaled single-cell whole-genome sequen