Society for Neuroscience Annual Meeting 2019

Visit the Sartorius Booth #1735 at SFN to learn the latest trends and applications in live-cell neuronal analysis, including our Neuronal Activity Application.

Developing the next generation of therapies for brain disease relies on advanced technologies that provide detailed insights into the morphology, function and activity of living cells.

IncuCyte S3® Live-Cell Analysis System for Neuroscience: enables real-time automated measurements of the dynamic changes and interactions in living neural cells directly from the incubator.

• Automated image acquisition and analysis of neuronal cell activity, cell health, structure and neuro-immune function in 96-and 384-well formats
• Visually verify morphological changes and validate measurements with images and movies
• Conduct week- or month-long studies on sensitive cells while they sit undisturbed in your incubator

CompacT SelecT™ is the leading automated cell culture system, used in labs across commercial and academic life science. It is now available with new options to enable the culturing of complex cell lines for oncology programs, stem cell research and toxicity studies.
 

At the show, our R&D scientists will be presenting a poster highlighting the power of live-cell analysis for quantification of neuro-pathology. Learn how IncuCyte® S3 Live-Cell Analysis System for Neuroscience and applications can enable new discoveries – all from inside your incubator!

Abstract Title: Improved Alzheimer's disease models using neuronal and microglial live-cell analysis in 2D and 3D

Authors: *S. L. ALCANTARA1, J. RAUCH2, G. LOVELL1, L. OUPICKA2, J. TRIGG1, A. OVERLAND2, M. BOWE2, N. HOLTZ2, E. ENDSLEY2, C. SCHRAMM2, D. APPLEDORN2, D. TREZISE1, T. DALE1;

1 BioA Applications, Sartorius (Essen Bioscience), Welwyn Garden City, United Kingdom;
2 BioA Applications, Sartorius (Essen Bioscience), Ann Arbor, MI

Session Number: 299

Session Title: Alzheimer's Disease: Neurotoxicity, Inflammation, and Neuroprotection

Date and Time: Monday Oct 21, 2019 8:00 AM - 12:00 PM

Location: McCormick Place Hall A

Abstract Control Number: 11199

Development of more translational in vitro Alzheimer’s disease (AD) models will benefit understanding of neuro-pathology.  To enable improved AD models, and to obtain a greater insight on the role of microglia in disease processes, we have developed techniques for long term measurements of cell health, morphology and function in 2D and 3D using live-cell analysis.

Healthy (hN6) and AD patient (hAD2) derived (Axol, Cambridge, UK) neuro-progenitor cells were seeded, differentiated, and matured for up to 90 days and monitored throughout using an IncuCyte® S3 for Neuroscience. After 14 d post differentiation in mono-culture, hAD2 iPSC-derived neurons yielded lower neurite outgrowth compared to the hN6 (neurite length of 80 ± 2.9 or 48 ± 2.6 mm/mm2 for hN6 or hAD2, respectively, mean ± SEM; 48 replicates). In a 3D spheroid model, culturing in ultra-low-attachment (ULA) plates, hAD2s demonstrated enhanced spheroid growth (63 ± 7 or 119 ± 4 % change for hN6 or hAD2, respectively, 2-5 replicates). The impact of AD related peptides (Tau and Aβ1-42) on neuronal health and function was quantified using nuclear labeled SH-SY5Y cells or rat primary cortical neurons in mono- or co-cultured with rat primary astrocytes. Tau and Aβ1-42 peptides induced a time- and concentration-dependent neuronal toxicity, with a reduction in neuronal number, neurite length and activity measured by calcium signalling.

Phagocytosis by microglia of pHrodo® labeled aggregated peptides was quantified as an increase in fluorescence following entry into acidic intracellular compartments.  Rapid engulfment of aggregated Aβ1-42 peptide was observed in mouse BV-2 cells (fluorescent area 18.1 ± 1.2 mm/mm2 at 48 h, 3 replicates).  Little or no engulfment was observed in undifferentiated SH-SY5Y cells or when the Aβ1-42 was not aggregated.  hiPSC-derived monocytes (Axol) were differentiated for 14 d into microglia. Addition of pHrodo® labelled aggregated Aβ1-42 (3.7 – 100 µg/ml) yielded concentration-dependent phagocytosis (maximum fluorescent area: 2.88x105 ± 0.02 µm2 at 24 h for 33 µg/ml peptide, 2 replicates). To elucidate the involvement of scavenger receptors in peptide uptake, CD204 and CD36 antibodies known to block scavenger receptors A and B, respectively, were added with aggregated Aβ1-42 or E.coli bio-particles®.  Whilst the CD204 antibody inhibited the uptake of both reagents, CD36 selectively attenuated Aβ1-42 peptide engulfment. 

Taken together, these data show that long term monitoring, alongside combining multiple readouts from advanced cellular models, has the potential to deliver greater biological insight into neurological disorders, therefore contributing to drug discovery.

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