A new benchtop system for simple and versatile introduction of macromolecules into human lymphocytes and CD34+ cells by microfluidic squeezing
Bibliographic record
Abstract
Abstract Intracellular delivery of molecules is a key step in biological research that enables the development of new methods of cell and gene-based therapies. Traditional methods for intracellular delivery are limited in a) delivering a wide variety of cargos, b) efficient delivery to a broad range of cell types, and c) preventing unwanted genetic dysregulation. To address these challenges, we have developed the CellPore™ system, a benchtop device for gentle, yet efficient delivery of cargos to a range of primary cell types. The CellPore™ system is a microfluidic platform that creates transient disruptions in the plasma membrane via mechanical deformation by applying pressure, thereby allowing cargo entry to the cytosol. We developed a simple and straightforward workflow to determine optimal delivery conditions. This workflow consists of cell isolation, cargo preparation for delivery across a range of system pressures, followed by viability and delivery efficiency measurements. Optimized parameters were subsequently applied to deliver eGFP and mCherry mRNA as well as B2M-targeting Cas9 ribonucleoproteins to human unactivated T and CD34+ cells. Importantly, edited cells were functionally active after manipulation by the CellPore™ system, and subsequent studies in T cells revealed minimal transcriptional perturbations compared to electroporated samples. These results demonstrate the importance of having a comprehensive understanding of the impact a delivery modality can have in a biological system. The CellPore™ system enables efficient delivery of cargos to target cells, while safeguarding key functional attributes. This simple workflow allows the CellPore™ system to be easily integrated into cell therapy research protocols.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".