Insphero GravityTRAP User manual

Product Manual
GravityTRAP™ ULA Plate
www.insphero.com
ISP-09-001
www.insphero.com

GravityTRAP™ ULA Plate Manual
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Contents
Introduction 3
GravityTRAP™ ULA Components 5
Generating 3D microtissues 8
Additional materials required 8
Preparation 9
Pre-wetting 9
Microtissue seeding 10
Sedimentation/Spheroid maturation 11
Medium exchange in the GravityTRAP™ ULA Plate 12
Analysis and assays in the GravityTRAP™ ULA Plate 13
Annex 1: Microtissue harvest from GravityTRAP™ ULA Plates 14
Annex 2: Troubleshooting guide 16
Annex 3: Step-by-step protocol for HCT116 & HEY microtissues 17
Version 2.0, July, 2015
451-0009-01-B

GravityTRAP™ ULA Plate Manual
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Introduction
The GravityTRAP™ Ultra-Low Attachment
(ULA) plate1represents a simple, exible,
and automation-compatible platform for the
generation, long-term cultivation, observation
and testing of scaold-free 3D microtissue
spheroids in 96-well format. Each plate
consists of a special non-adhesively coated
96-well, sterile-packaged GravityTRAP™ ULA Plate and lid.
InSphero recommends GravityTRAP™ ULA plates for the generation of
spheroids using immortalized or modied cell lines that are known to
readily form microtissues, or as a starting point for investigating whether or
not a cell line can form self-aggregating, scaold-free spheroids. InSphero
recommends our patented GravityPLUS™ Hanging Drop System (ISP-06-
001, ISP-06-010) if generating spheroids in more complex 3D cell culture
scenarios, such as when using primary cells, cell lines that are sensitive to
self-assembly, or when generating co-culture microtissues (e.g., tumor/
stroma). In such cases, the GravityPLUS™ Hanging Drop System provides
the greatest opportunity for success.
1 The GravityTRAP™ ULA Plate and GravityPLUS™ Plate and related technology are protected by several granted and pending patents
world-wide.

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Figure 1: Spheroid formation in the GravityTRAP™ ULA Plate begins with initial seeding
of cells in suspension, followed by a brief spin to concentrate cells. Following microtissue
maturation,the SureXchange™ ledge of the tapered well facilitates medium exchange and
compound dosing without disturbing or losing the microtissue.
Advantages of the GravityTRAP™ ULA Plate:
1. Convenient scaold-free formation of spheroids via cellular self-assembly in
ultra-low attachment (ULA-treated) plates
2. SureXchange™ tapered ledge and culture chamber facilitates easy medium
exchange and prevents microtissue loss during long-term spheroid growth and
analysis
3. 1 mm diameter at bottom observation chamber enables simple spheroid
observation, and greater working eld-to-eld distance reduces well-to-well
imaging cross-talk compared to standard 96-well plates
4. 3D-optimized protocols available for analysis in GravityTRAP™ ULA Plate

GravityTRAP™ ULA Plate Manual
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GravityTRAP™ ULA Plate Components
The complete GravityTRAP™ ULA Plate assembly consists of the following
components:
1. Bottom GravityTRAP™ ULA Plate (96-well) (A)
2. Lid (B)
Figure 2:
Components of
the GravityTRAP™
ULA Plate