Cell Culture
Figure 5: Where 3D cell culture is expected to impact most
Basic research – cell culture methodology Tissue/organ engineering
Drug discovery – safety assessment /toxicology Drug discovery – secondary screening/lead opt. Drug discovery – target validation Basic research – developmental biology Basic research – cell biology
Drug discovery – primary screening Clinical research
Cell supply & cryopreservation Drug discovery – DMPK
2.84
1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 MEAN RATING SCALE 1 to 5, where 1 = no impact and 5 = major impact
© HTStec 2010 3.22 3.09 4.46
3.91 3.93 3.96
3.72
3.52 3.54
3.32
Figure 6: Assay types successfully used/ investigated in 3D cell culture
Metabolic activity assay Cell signalling assay Cell migration Cell proliferation Cell viability
Electrical recording of field potentials Hepatocyte metabolism Reporter gene assay Chemotaxis
Patch clamping 23% 18% 12% 7% 6% © HTStec 2010
0% 10% 20% 30% 40% 50% 60% 70% % Successfully used/investigated
63% 61% 52% 31% 29%
recently been chosen by The National Institute of Standard Technology (NIST) as the reference 3D tissue engineering scaffold. The non-degradable 3D Insert™-PS has been developed for conducting routine 3D cell culture. Made from the same mate- rial as traditional tissue culture plates, 3D Biotek has essentially engineered 2D polystyrene into a 3D scaffold. Combining the precisely engineered 3D structure with the inherent transparency of polystyrene material, creates an ideal 3D Insert™- PS scaffold for performing easy 3D cell culture. These PS scaffolds are compatible with most 2D assays and allow researchers to monitor 3D cell growth by simply using an inverted light micro- scope. 3D Biotek has initiated a partnership with BioCellChallenge SAS, a drug delivery company in France, and developed the world’s first 3D cell transfection kit, which combines 3D Insert™-PS and Transfection Reagent into one kit. This is the world’s first 3D in vitro transfection technology, which now allows researchers to achieve high delivery efficiencies of plasmid DNA into 3D cul- tured cells (Figure 8).
BellBrook Labs (
www.bellbrooklabs.com) has developed a device and methods to address the need for automatable and high-content assays for
Figure 7: Most important objectives of transitioning from 2D to 3D culture
Better quality of assay results Enhanced cell viability/responsiveness Greater biological relevance
Simpler automation Labour/cost savings Higher throughput Higher yield
3.37 3.24 2.79 2.56
1.00 2.00 3.00 4.00 5.00 6.00 7.00 MEAN RANKED ORDER, where 1 = least important and 7 = most important
© HTStec 2010
Figure 9: A) Schematic of BellBrook Labs IUVO MC5250 Microchannel. B&C) BxPC3 cells suspended in 3D Type I Collagen were incubated 18 hours and then stained for EdU incorporation using Invitrogen ClickIT EdU Imaging Kit. Hoescht staining of nuclei is shown in blue, and cells with incorporated EdU are shown in red. An average of 19% of untreated cells (B) have incorporated EdU, compared to 0% for cells treated (C) with 10µM Cycloheximide. n = 4, p
28 Drug Discovery World Summer 2010 6.50 5.20 5.05
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