AI Article Synopsis

  • Automated high-throughput methods are essential for tracking mammalian cell growth to improve cell line characterization and identify genetic components for cell engineering.
  • The described noninvasive assay uses plate reader measurements of the pH indicator phenol red to create a cell growth index that correlates with actual cell counts.
  • This method is effective for analyzing both suspension and adherent cell lines under varying conditions, enhancing the study of engineered cell lines and helping identify desired genetic promoters through fluorescence.

Article Abstract

Automated high-throughput methods that support tracking of mammalian cell growth are currently needed to advance cell line characterization and identification of desired genetic components required for cell engineering. Here, we describe a high-throughput noninvasive assay based on plate reader measurements. The assay relies on the change in absorbance of the pH indicator phenol red. We show that its basic and acidic absorbance profiles can be converted into a cell growth index consistent with cell count profiles, and that, by adopting a computational pipeline and calibration measurements, it is possible to identify a conversion that enables prediction of cell numbers from plate measurements alone. The assay is suitable for growth characterization of both suspension and adherent cell lines when these are grown under different environmental conditions and treated with chemotherapeutic drugs. The method also supports characterization of stably engineered cell lines and identification of desired promoters based on fluorescence output.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-4063-0_5DOI Listing

Publication Analysis

Top Keywords

cell growth
12
cell
9
identification desired
8
measurements assay
8
cell lines
8
simultaneous plate-reader
4
characterization
4
plate-reader characterization
4
characterization promoter
4
promoter activity
4

Similar Publications

Epstein-Barr nuclear antigen 1 (EBNA1), a sequence-specific DNA binding protein of Epstein-Barr virus (EBV), is essential for viral genome replication and maintenance and is therefore an attractive target for the therapeutic intervention of EBV-associated cancers. Several EBNA1-specific inhibitors have demonstrated the ability to block EBNA1 function in vitro, but practical delivery strategies for these inhibitors in vivo are still lacking. Here, we report an intelligent hierarchical targeting theranostic nanosystem (denoted as mZGOCS@MnO-P5) that integrates an azide (N3) terminal dual-targeting peptide (N3-P5), a tumor microenvironment-responsive degradable MnO nanosheet, and a mesoporous ZnGaO:Cr, Sn near-infrared persistent luminescence (NIR-PL) nanosphere (mZGOCS).

View Article and Find Full Text PDF

Mitochondrial DNA (mtDNA) is highly polymorphic, and host mtDNA variation has been associated with altered cancer severity. To determine the basis of this mtDNA-cancer association, we analyzed conplastic mice with the C57BL/6J (B6) nucleus but two naturally occurring mtDNA lineages, and , where mitochondria generate more oxidative phosphorylation (OXPHOS)-derived reactive oxygen species (mROS). In a cardiac transplant model, Foxp3+ T regulatory (Treg) cells supported long-term allograft survival, whereas Treg cells failed to suppress host T effector (Teff) cells, leading to acute rejection.

View Article and Find Full Text PDF

Using maize plants expressing an apoplast targeted Aspergillus niger ferulic acid esterase (FAEA), with FAEA driven by a Lolium multiflorum senescence enhanced promoter (LmSee1), we extended measurements of FAEA activity to late-stage senescing plants and measured the stability of FAEA activity following stover storage. The impact of FAEA expression on cell wall hydroxycinnamic acid levels and arabinoxylan (AX) cross-links, and on the levels of cell wall sugars, acetyl bromide lignin and sugar release following saccharification by a cocktail of cellulases and xylanases, was assessed during plant development to full leaf senescence. These were determined in both individual internodes and in combined leaves and combined internodes of FAEA expressing and control partner plants.

View Article and Find Full Text PDF

The mechanisms underlying the establishment of asymmetric structures during development remain elusive. The wing of Drosophila is asymmetric along the Anterior-Posterior (AP) axis, but the developmental origins of this asymmetry is unknown. Here, we investigate the contribution of cell recruitment, a process that drives cell fate differentiation in the Drosophila wing disc, to the asymmetric shape and pattern of the adult wing.

View Article and Find Full Text PDF

Aim: To explore the role of the hub gene Transforming Growth Factor Beta Induced (TGFBI) in Intervertebral disc degeneration (IDD) pathogenesis and its regulatory relationship with Membrane Associated Ring-CH-Type Finger 8 (MARCHF8).

Background: IDD is a prevalent musculoskeletal disorder leading to spinal pathology. Despite its ubiquity and impact, effective therapeutic strategies remain to be explored.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!