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Defined Bioscience

Application Notes and Posters

Application Notes

Defined Bioscience

High-density pluripotent stem cell expansion in a single-use bioreactor using HiDef S8

This study demonstrates that HiDef® S8, with Ready-CEPT™, supports rapid, high yield, carrier-free expansion of hPSCs in a stirred tank suspension bioreactor. In just five days, seven billion cells were generated in a two-liter BIOne single use bioreactor system operating at half-capacity, while maintaining high viability and expression of key pluripotency markers. The expanded hPSCs readily re-attached in 2D adherent culture, maintained normal morphology, and retained pluripotency after multiple passages and freeze thaw cycles. 

HiDef-S8 was specifically developed to meet the challenges of large-scale suspension culture and builds on the proven performance of HiDef® B8. Its defined, animal-free composition provides a reliable and scalable platform for seamlessly transitioning hPSC workflows from conventional monolayer cultures to high efficiency bioreactor systems. By eliminating the need for microcarriers and minimizing process complexity, HiDef-S8 reduces cost per cell, improves yield per unit of media, and supports the quality requirements of both research and translational applications.

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Optimized Expansion, Cryopreservation, and Recovery of Clinical Grade, PluriBank iPSCs Using a Defined Reagent System Kit

This study demonstrates that HiDef® B8, in combination with Ready-CEPT™ and PluriFreeze, enables high-efficiency, reproducible expansion and cryopreservation of clinical-grade human iPSCs (PluriBank™) under fully defined, xeno-free conditions. Two independent lines cultured in HiDef-B8 maintained high viability (>90%), robust proliferation (≥8-fold expansion per passage), and stable expression of pluripotency markers (OCT4⁺ SOX2⁺ SSEA4⁺) across multiple passages. Cryopreserved cells recovered with >80% viability and retained genomic integrity and pluripotency post-thaw. A reduced feeding schedule with HiDef-B8 preserved performance while lowering reagent use and handling time. These results validate a reliable, scalable, and cost-effective workflow for hiPSC manufacturing using chemically defined reagents. The combined system supports consistent performance for both routine culture and clinical translation, enabling standardized cell banking with reduced variability and improved operational efficiency.

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Comparative Analysis of 2D and 3D Differentiation Approaches for iPSC-Derived Neural Cells

The differentiation of induced pluripotent stem cells (iPSCs) into neural progenitor cells (NPCs) and neurons provides essential tools for understanding neural development, modeling diseases, and developing therapeutics. Protocols leveraging 2D monolayer cultures and 3D systems each offer distinct benefits, suiting diverse research objectives. While 2D methods are well-established for their simplicity and planar structure that has advantages for imaging, 3D approaches allow for the recapitulation of more physiologically relevant cellular microenvironments and cell types.

Defined Bioscience’s HiDef® B8 medium and reagents play an important role in the success of these workflows, ensuring consistent and reproducible iPSC expansion as a foundation for these downstream applications. This collaboration highlights the utility of iPSC technology in addressing some of the unmet needs in neuroscience. Contact Defined Bioscience to learn about related HiDef® B6 medium, HiDef® N2 neural supplement, thermostable FGF2-G3, and Ready-CEPT™ cocktail to support clonal robustness in single-cell expansion and reduce selection pressure for high passage and suspension PSC workflows.

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Defined Bioscience

Isolating Induced Pluripotent Stem Cells by Using the WOLF G2 to Sort Stemness-Related Surface Markers

Gentle cell sorting technologies are advantageous in the isolation and expansion of homogeneous hiPSCs, as is the use of fully defined and animal-free cell media. In collaboration with NanoCellect Biomedical, we grew hiPSCs in HiDef® B8 and used the WOLF G2 to identify and enrich for hiPSCs expressing SSEA-4 and TRA-1-60-R. HiDef-B8 grown hiPSCs showed healthy morphology, viability, and expression of these markers, with robust post-sorting recovery in the same medium.

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Defined Inputs. Consistent Outcomes
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Posters

Defined Bioscience
Varied growth factor performance across homologs

Growth factor production for enhanced growth of fish multipotent stem cells for cell-cultivated meat

Growth factor homologs across species can exhibit naturally evolved differences in function and in vitro stability. Continuing work previously presented at ISSCR, we studied changes in thermostability of recombinantly produced FGF2 homologs in advance of cell culture efforts. This work highlighted the value of variations in nature that can contribute to growth factor performance. Presented at ISSCR 2024 - Cincinnati Symposium.

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FGF2 homologs in the context of cell culture

FGF2, part two: evaluation of homologs for use in human and animal cell culture for the life sciences and cultivated meat

Sequence homologs of Basic Fibroblast Growth Factor (bFGF or FGF2) have exhibited differential performance in cell growth across species, with clear implications for improving cost and performance of human- and animal-derived cell culture. Continuing from work presented at ISSCR 2022 (see previous poster below), we evaluated several candidates for proliferation of human- and animal-derived cell lines. These results highlight the value of growth factor homolog screening when optimizing cell culture conditions. Presented at ISSCR 2023.

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Defined Bioscience
HiDef® B8 for single-cell clonal expansion

Defined cGMP animal-origin-free medium supports PSC during, and promotes single-cell clonal expansion after, microfluidic sorting

Fully defined and animal/serum-free media, manufactured under strict cGMP protocols and with traceable sourcing, are of high value in pluripotent stem cell isolation and expansion. Continuing from our collaboration with NanoCellect Biomedical (see application note below), we explored the value of our such medium, HiDef® B8, in the culture of hiPSCs. Presented at ISSCR 2023.

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HiDef® B8 optimization using DOE

Optimized HiDef® B8 Reduces Costs and Improves Accessibility for PSC Research

Consistency in growth media remains a challenge for stem cell research and manufacturing. Continuing from our work presented at ISSCR 2022 (see poster below), we continued our validation of HiDef® B8 using DOE-driven approaches, focusing on three components of the medium. Of the variants tested, HiDef-B8 continued to outperform based on function and cost. Presented at ISSCR 2023.

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Defined Bioscience
FGF2 variants for life science and cultivated meat

Development of FGF2 homologs for applications in human stem cell culture and the cultivated meat industry

Naturally occurring isoforms of growth factors offer a range of unique features across diverse applications, and suggest means for evolution-driven improvements in stability, function, and efficacy. We produced a range of FGF2 homologs, coupled with the structure determination of thermostable human variant FGF2-G3, to inform the development of new growth factor solutions. Presented at ISSCR 2022.

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Engineering approaches to medium optimization

Defining Culture: Cost-effective Medium to Improve Stem Cell Reproducibility

Current growth medium solutions for the cell culture space are oftentimes inconsistent, variable, and costly, warranting the development of more optimized alternatives. We applied a design of experiments (DOE) to fit a response surface methodology (RSM) model to HiDef® B8 medium for continued optimization in stem cell culture. Presented at ISSCR 2022.

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FGF2-G3 usage in HiDef® B8

Growth factor production and optimization for fully-defined media in induced pluripotent stem cell culture

Serum-containing media present major obstacles for the cell culture space, including batch-to-batch variability, undefined componentry, and the introduction of growth-inhibiting factors. We produced FGF2-G3, a thermostable variant of human FGF2, and studied its use in the context of HiDef® B8, a serum-free, fully-defined formulation for the culture of stem cells. Presented at ISSCR 2022.



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Turning innovation into impact

Applications

See how Defined Bioscience products support research in disease modeling, drug discovery, and beyond.
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Disease Modeling

Disease Modeling Using Human Induced Pluripotent STEM Cells (iPSCs)

Defined Bioscience
Drug Discovery and Toxicology

IPSC-Based Models Transforming How Drugs are Tested for Safety and Effectiveness.
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Regenerative Medicine

Regenerative Medicine Using Human Pluripotent STEM Cells (iPSCs)
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Immuno-oncology

Immuno-oncology and Immunotherapy Using Human Induced Pluripotent STEM Cells (iPSCs)

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