Biology subjects

Sramkova, Z.

Publications and source records attributed to Sramkova, Z..

3 recordsLinked to original sources

bioRxiv2024

Enhancing reproducibility and decentralization in single cell research with biocytometry

Biomedicine today is experiencing a shift towards decentralized data collection, which promises enhanced reproducibility and collaboration across diverse laboratory environments. This inter-laboratory study evaluates the performance of biocytometry, a method utilizing engineered bioparticles for enumerating cells based on their surface antigen patterns. In a decentralized framework, spanning 78 assays conducted by 30 users across 12 distinct laboratories, biocytometry consistently demonstrated significant statistical power in discriminating numbers of target cells at varying concentrations as low as 1 cell per 100,000 background cells. User skill levels varied from expert to beginner capturing a range of proficiencies. Measurement was performed in a decentralized environment without any instrument cross-calibration or advanced user training outside of a basic instruction manual. The results affirm biocytometry to be a viable solution for immunophenotyping applications demanding sensitivity as well as scalability and reproducibility and paves the way for decentralized analysis of rare cells in heterogeneous samples.

cell biology↗

bioRxiv2024

Massively parallel identification of single-cell immunophenotypes

Translating insights from single-cell analysis into actionable indicators of health and disease requires large-scale confirmatory studies. We introduce biocytometry, a novel method utilizing engineered bioparticles for multiparametric immunophenotyping in suspension, enabling simultaneous measurement across thousands of assays with single-cell sensitivity and a wide dynamic range (1 to 1,000 target cells/sample). The technical validation of biocytometry revealed strong alignment with established technologies (mean bias = 0.25%, LoA = -1.83% to 2.33%) for low-sensitivity settings. Biocytometry excelled in high-sensitivity settings, consistently showcasing superior sensitivity and specificity (LoB = 0), irrespective of the sample type. By employing multiparametric target cell identification, we harnessed the homogeneous assay workflow to discern cell-specific apoptosis in mixed cell cultures. Potential applications include monitoring rare premalignant subpopulations in indications such as smoldering multiple myeloma (SMM), enhancing the detection of circulating tumor cells (CTCs), advancing pharmacokinetic assessments in chimeric antigen receptor (CAR) T-cell therapies, and improving the accuracy of minimal residual disease (MRD) evaluations. Additionally, the high throughput and cell-specific readout capabilities might provide substantial value in drug development, especially for the analysis of complex sample matrices, such as primary cell cultures and organoids.

synthetic biology↗

bioRxiv2020

miR-183/96/182 cluster is an important morphogenetic factor targeting PAX6 expression in differentiating human retinal organoids

MicroRNAs (miRNAs), a class of small, non-coding RNA molecules represent important regulators of gene expression. Recent reports have implicated their role in the cell specification process acting as "fine-tuners" to ensure the precise gene expression at the specific stage of cell differentiation. Here we used retinal organoids differentiated from human pluripotent stem cells (hPSCs) as a model to closely investigate the role of a sensory organ-specific and evolutionary conserved miR-183/96/182 cluster. Using a miRNA tough decoy approach, we inhibited the miR-183/96/182 cluster in hPSCs. Inhibition of the miRNA cluster resulted in an increased expansion of neuroepithelium leading to abnormal "bulged" neural retina in organoids, associated with upregulation of neural-specific and retinal-specific genes. Importantly, we identified PAX6, a well-known essential gene in neuroectoderm specification, as a target of the miR-183/96/182 cluster members. Taken together, the miR-183/96/182 cluster not only represents an important regulator of PAX6 expression, but it also plays a crucial role in retinal tissue morphogenesis.

molecular biology↗