Comparative Analysis of 2D and 3D Differentiation Approaches for iPSC-Derived Neural Cells
Protocol for creating neural cells from iPSCs in collaboration with Novoron
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.
Transitioning to fully defined HiDef-B8 for adherent human pluripotent stem cell cultures
Simple transition to HiDef-B8 from other growth media
HiDef-B8 is a low-cost, animal-free, and weekend-free growth medium for the culture of pluripotent stem cells (PSCs), focusing on long-term expansion capacity and maintenance of a stable, undifferentiated state. This guide aids users in easily transitioning from other medium types, including mTeSR-Plus, to HiDef-B8.
Growth factor production for enhanced growth of fish multipotent stem cells for cell-cultivated meat
Varied growth factor performance across homologs
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.
FGF2 homologs in the context of cell culture
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.
HiDef-B8 for single-cell clonal expansion
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.
Optimized HiDef-B8 Reduces Costs and Improves Accessibility for PSC Research
HiDef-B8 optimization using DOE
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.
Gentle sorting of hiPSCs grown in HiDef-B8 medium
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.
FGF2 variants for life science and cultivated meat
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.
Defining Culture: Cost-effective Medium to Improve Stem Cell Reproducibility
Engineering approaches to medium optimization
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.
FGF2-G3 usage in HiDef-B8
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.
