Abstract Thu035: Growth Factor-Based Manufacturing of Human Pluripotent Stem Cell-Derived Cardiomyocytes Using the Vertical Wheel Bioreactor System
Bibliographic record
Abstract
Background: Although several methods have been reported for the large-scale generation of hPSC-derived cardiomyocytes (hPSC-CMs) in bioreactors exclusively using small molecules, no comparable bioreactor-based protocol has been established using growth factors, despite evidence that differentiation with optimized concentrations of BMP4, Activin A, and bFGF offers finer control of cardiomyocyte phenotype. Here, we develop scalable hPSC expansion and growth factor based cardiac differentiation protocols using the Vertical Wheel Bioreactor (VWBR) system. Methods and Results: The expansion of undifferentiated hPSCs in VWBRs was optimized by varying parameters including cell inoculation density, agitation rate, and media exchange schedule. Next, we examined the impact that different hPSC culture conditions had on subsequent cardiac differentiation potential. Notably, hPSCs expanded at different densities and agitation rates all yielded high purity hPSC-CMs when differentiated in either 6-well plates or VWBRs. However, hPSCs initially expanded in VWBRs showed significantly lower cardiomyocyte yields than hPSCs expanded in 2D culture. Examination of differentiation kinetics revealed that 3D expanded hPSCs exhibited accelerated mesoderm commitment relative to their 2D counterparts, as well as a significant fraction of endoderm contamination by as early as day 3 of differentiation. Modification of our aggregation and mesoderm induction steps greatly improved cardiogenesis and enabled reliable production of >1x10 6 /mL cardiac troponin T + (cTnT) hPSC-CMs expressing MLC2v. Highlighting the versatility of our protocol, simple changes in the BMP4:Activin A ratio enabled second heart field-based differentiation and production of atrial-like cardiomyocytes in VWBRs. Lastly, we show that our developed expansion and differentiation protocols were reproducible and economic in 500mL VWBRs, enabling the production of 1.1x10 6 hPSC-CMs/mL at a mean purity of 93% cTnT + . Conclusions: This novel bioreactor-based protocol will enable cardiac cell therapy and tissue engineering applications by providing scalable and consistent production of hPSC-derived cardiac cell products.
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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".