Single-Cell RNA Sequencing in Ovarian Cancer: Current Progress and Future Prospects

Progress in Biophysics and Molecular Biology, Год журнала: 2025, Номер unknown

Опубликована: Янв. 1, 2025

Язык: Английский

Isolation of planarian viable cells using fluorescence‐activated cell sorting for advancing single‐cell transcriptome analysis

Genes to Cells, Год журнала: 2023, Номер 28(11), С. 800 - 810

Опубликована: Сен. 18, 2023

Preparing viable single cells is critical for conducting single-cell RNA sequencing (scRNA-seq) because the presence of ambient from dead or damaged can interfere with data analysis. Here, we developed a method isolating adult planarian bodies using fluorescence-activated cell sorting (FACS). This was then applied to both pluripotent stem (aPSCs) and differentiating/differentiated cells. Initially, employed violet instead ultraviolet (UV) laser excite Hoechst 33342 reduce cellular damage. After optimization staining conditions FACS compensation, generated profiles similar those created previous that UV laser. Despite successfully obtaining high-quality aPSCs, non-aPSCs produced low-quality reads (i.e., <60% possessing barcoding mRNAs). Subsequently, identified an effective gating condition excluded tissue debris without staining. non-staining isolation strategy not only reduced post-dissociation time but also enabled scRNA-seq results all types >80%). Taken together, these findings imply may be beneficial planarians other organisms tissues studies.

Язык: Английский

Single Cell RNA Sequencing and Data Analysis

Elsevier eBooks, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Язык: Английский