Cultrex® Embryoid Body Formation Kit
The Cultrex® Embryoid Body Formation Kit offers a useful tool for generating consistent, reproducible and identically-sized embryoid bodies (EBs). The kit has been qualified to form EBs from human pluripotent stem cells (hESCs and iPSCs).
Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have the unique ability to differentiate into cells of three embryonic germ layers (endoderm, ectoderm and mesoderm).
The majority of differentiation protocols start with the generation of embryoid bodies (EBs) - three-dimensional multicellular aggregates of pluripotent stem cells.
Although pluripotent stem cells placed in suspension culture will form EBs spontaneously, the efficiency of this process varies and is not reproducible. Most current methods avoid preparing single cell suspension,
due to low survival of pluripotent stem cells in single cell culture, and require keeping cells in large clumps, thus generating EBs of different size and shapes.
Size of EBs is one of the key factors affecting differentiation of PSCs and needs to be controlled.
To solve these problems and help to improve reproducibility of differentiation procedures, we have developed the Cultrex® Embryoid Body Formation Kit, comprising:
- Specialized EB Formation Matrix - to stimulate cell aggregation and development of EBs
- ROCK Inhibitor Y-27632- which significantly increases survival of PSCs in single cell culture
- 96 Well EB Formation Low-Adhesion Plate that allows researchers to control the size of EBs and create a single EB in each well
Figure 1. Cultrex® Embryoid Body Formation Kit helps to generate identical in size EBs. A, B, C - EBs formed from human iPSCs using Cultrex® Embryoid Body Formation Kit; shown EBs generated with 3,000 cells per well after 48 hours.
D, E, F - EBs formed from human iPSCs using conventional method; 3,000 cells per well after 48 hours.
Each image (A-F) represents individual well of 96-well plate.
Figure 2. EBs generated from human iPS cells using Cultrex® Embryoid Body Formation Kit can be differentiated in three germ lineages.
EBs were formed from iPSCs using Cultrex® Embryoid Body Formation Kit and incubated for 21 days in DMEM-10%FBS in 96-well EB Formation Plate followed by quantitative RT-PCR analysis.
Undifferentiated iPS cells were cultured in mTeSR1 medium on RGF BME-coated plate.
Organoid Growth Matrix
3D Triculture Model
Physiologically Relevant Cell Culture
Cultrex® is a Trevigen trademark