3D Human iPSC Line (Control)
Applied StemCell Inc. provides high quality, 3D cultured induced pluripotent stem cells (3D iPSCs) grown from Applied StemCell’s well-characterized, integration-free control iPSC line (ASE-9203). These 3D iPSCs were cultured in a peptide nanofibril 3D matrix, MyEZGel™ 3D-iPSC Matrix (ASR-3053/3054) using xeno-free conditions. The MyEZGel™ 3D-iPSC matrix mimics an in vivo-like microenvironment resulting in spheroidal iPSCs that are more robust than 2D cultured iPSCs, and have longer pluripotency and better differentiation potential as determined by in vivo teratoma assays.
These ready-to-use ASE-9050 3D-iPSCs can be thawed and expanded directly into the MyEZGel™ 3D-iPSC Matrix (ASR-3053) provided along with the cells using easy-to-use protocols. These 3D-iPSCs can be used for further in vitro differentiation into a somatic lineage of choice, as well as used as a control line for experiments using your 3D cultured patient-derived or engineered.
Applied StemCell, Inc. also provides custom 3D- iPSC culture and expansion services for customer provided iPSC lines. For further information, please contact us for more details.
3D iPSCs Cultured in MyEZGel™ 3D-iPSC Matrix
Figure 1. Optimal seeding density and cell viability assay after cyropreservation. Control Human iPSC Line was seeded onto MyEZGel™ 3D-iPSC Matrix in mTeSR1 media at three different seeding densities, 1, 3 and 5 x 10^5 cells/ well. The cells were cultured for 4 days and total fold expansion determined. Cell seeded at the 1x10^5 cells/well showed the highest expansion after culturing. These observations suggest that maximum expansion occurs using low seeding densities. Cyropreserved 3D iPSCs were thawed in 3D matrix at 3 seeding densities and viable cells were counted. All three seeding densitites showed 80% cell viability.
Figure 2. In situ Immunohistochemistry for pluripotency markers. Control human 3D iPSCs (ASE-9050) at passage 3 (p3) were stained for pluripotency marker OCT3/4 directly in the 3D matrix on day 5 of culture. The 3D iPSCs showed high level expression of pluripotency marker.
Figure 3. Control Human 3D iPSCs culutred in MyEZGel™ 3D-iPSC Matrix show expression of key pluripotency markers.
Figure 4. Teratoma formation comparing 2D (passage #21) and 3D (passage #34) cultured hiPSCs show better in vivo differentation into germ layer by 3D iPSCs as compared to 2D hiPSCs. In fact, 3D iPSCs at a later passage (p34) were more pluripotent and showed better differentiation potential compared to 2D hiPSCs at an earlier passage (p21).
- Li, L., & LaBarbera, D. V. (2017). 3D High-Content Screening of Organoids for Drug Discovery. Reference Module in Chemistry, Molecular Sciences and Chemical Engineering. 388-415. doi.org/10.1016/B978-0-12-409547-2.12329-7