Protocols

iPSC reprogramming from human peripheral blood using Sendai Virus mediated gene transfer

Wenli Yang, Jason A. Mills, Spencer Sullivan, Ying Liu, Deborah L. French and Paul Gadue

This protocol allows efficient generation of integration-free iPS cells from a small amount of peripheral blood. Peripheral blood mononuclear cells (PBMCs) are cultured to expand the erythroblast (EB) population. They are then used to derive iPS cells using four recombinant Sendai viral vectors, expressing the four reprogramming factors Oct4, Sox2, Kfl4 and c-Myc.

Robust generation of hepatocyte-like cells from human embryonic stem cell populations

C.N. Medine, B. Lucendo-Villarin, W. Zhou, C.C., West, D.C. Hay

Despite progress in modelling human drug toxicity, many compounds fail during clinical trials due to unpredicted side effects. The cost of clinical studies are substantial, therefore it is essential that more predictive toxicology screens are developed and deployed early on in drug development.

Monolayer endoderm differentiation from human ESCs

Cheng X., Ying L., Lu L., Galvao A.M., Mills J.A., Lin H.C., Kotton D.N., Shen S.S., Nostro M.C., Choi J.K., Weiss M.J., French D.L., Gadue P.

When embryonic stem cells are differentiated in the presence of activin A in serum-free conditions, an endoderm progenitor population defined by the coexpression of either Brachyury, Foxa2 and c-Kit, or c-Kit and Cxcr4 is generated. Specification of these progenitors with bone morphogenetic protein-4 in combination...

Blood - SeV derived fibroblast generated iPSCs

Laurence Daheron and Sunita D'Souza

CytoTune™-iPS Reprogramming System uses vectors based on replication in competent Sendai virus (SeV) to safely and effectively deliver and express key genetic factors necessary for reprogramming somatic cells into iPSCs. In contrast to many available protocols, which rely on viral vectors that integrate into the genome of the host cell...

Protocol for directed differentiation of human pluripotent stem cells toward a hepatocyte fate

Jun Cai, Ann DeLaForest, Joseph Fisher, Amanda Urick, Thomas Wagner, Kirk Twaroski, Max Cayo, Masato Nagaoka, Stephen A Duncan

The directed differentiation of human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) into hepatocytes could facilitate a rational study of the molecular mechanisms underlying human liver development as well as provide a renewable source of exogenous hepatocytes for drug toxicity testing and cell-based therapeutics.