Reseeding Decellularized Porcine Ventricular Extracellular Matrix with Isolated Primary Human Atrial Cells
Bibliographic record
Abstract
As a result of pioneering research unveiling the influence of extracellular environment in determining stem/progenitor cell differentiation, a myriad of cellular scaffolds have been developed in order to optimize the delivery of these cells in a diseased environment as a technique to improve their efficacy in stem cell therapy. Although a variety of scaffolds are being studied in different contexts, the most physiologically relevant ones are those that most closely resemble the environment to be treated. Our lab has a keen interest in cardiovascular disease and has recently optimized a technique that decellularizes and reseeds isolated sections of healthy porcine ventricular tissue with primary human atrial cells. Decellularization involved a multi‐step, multi‐reagent washing process over a period of four days, after which confirmation of a lack of cells was determined by DAPI staining and subsequent western blotting looking for the absence of β‐tubulin. After decellularization, our objective was to successfully seed the decellularized tissue slices via co‐incubation of isolated primary human atrial fibroblasts (hAFs) or human c‐kit (+) cardiac progenitor cells (hCCs). These two cell types were chosen to compare how hAFs and hCCs respond differently to healthy extracellular matrix (ECM). Cells were isolated from the atrial appendage of patients undergoing open heart surgery by mincing followed by collagenase digestion and subsequent isolation of hCCs was performed using a c‐kit (+) magnetic bead isolation procedure. HAFs and hCCs were incubated separately on sections of decellularized porcine ventricular ECM over a period of 24 hours and then visualized using immunofluoresence. Collagen‐1, fibronectin, and versican antibodies were used to image the ECM structure and DAPI was used to identify cells embedded in the recellularized matrix. Successful seeding of cells was also evaluated by western blot detection of β‐tubulin in samples not prepared for immunofluorescence. In conclusion, our technique was successful in reseeding decellularized porcine ventricular extracellular matrix and can be extrapolated to detect other proteins in these tissues without having to reseed an entire decellularized heart. This can be used to compare differences among cell types and their responses between healthy and damaged or diseased cardiac ECM among different hearts or even within the same heart. Support or Funding Information Mazankowski Alberta Heart Institute University of Alberta Hospital Foundation Alberta Innovates Health Solutions
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| 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) | 0.001 | 0.001 |
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 source (direct Gemma or distilled Codex), 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".