Stem Cell Protocols
Below are links to a number of different online protocols for the derivation and culture of human ES and iPS cells. In addition to these protocols, we highly recommend StemBook, which is publishing a growing list of methods to generate induced pluripotent stem cells and differentiate them into various lineages.
Derivation of Human Embryonic Stem Cell Lines
PDF of Chapter 4 from “Human Embryonic Stem Cells: The Practical Handbook” (2007, Wiley Publishing) describes methods used by Harvard University researchers to establish human ES cell lines.
Optimized protocol for derivation of human embryonic stem cell lines.
Camarasa MV, Galvez VM, Brison DR, Bachiller D. In: Stem Cell Reviews and Reports September 2012, Volume 8, Issue 3, pp 1011-1020
|Human iPS Cell Derivation/Reprogramming||Daley Lab, Children’s Hospital, Boston||Retrovirus|
|Generation of Human induced Pluripotent Stem Cells||Center for iPS Cell Research and Application, Kyoto University||Retrovirus|
|Human iPS Cell Generation with hSTEMCCA||Center for Regenerative Medicine, Boston University||Lentivirus, STEMCCA|
|Human fibroblast reprogramming: Lentiviral method (hSTEMCCA)||iPS Core Facility, Harvard University||Lentivirus, STEMCCA|
|Reprogramming human fibroblasts using an episomal vector system||iPS Core Facility, Harvard University||Episomal|
|Reprogramming Fibroblasts with Sendai||Mount Sinai School of Medicine||Sendai Virus|
|iPSC Reprogramming From Human Peripheral Blood Using Sendai Virus Mediated Gene Transfer||University of Pennsylvania||Sendai Virus|
Basic Pluripotent Stem Cell Culture Protocol
StemBook protocol from the Unversity of Massachusetts Medical School. Covers reagents and techniques required to grow, passage, and cryopreserve human pluripotent stem cells on mouse feeders.
Standard Operating Procedures (SOP) for the culture of human iPS
Culture protocols from the Harvard iPS Cell Core Facility
Detailed set of PDFs with figures covering MEF feeder derivation and culture and the thawing, splitting, selection and maintenance of human ES cells.
This PDF (2004) covers topics such as: derivation, growth and use of MEFs, bulk and microdissection passaging of human ES cellsand preparation of human ES cellsfor karyotyping.
A collection of methods by a team ofstem cell core facility heads. The protocols span the range of diverse techniques inherent to the interdisciplinary nature of stem cell research.
NIH Center for Regenerative Medicine Protocols
Protocols for the generation, characterization and differentiation of iPS cells gathered by the CRM.
Human Stem Cell Technology and Biology: A Research Guide and Laboratory Manual
An authoritative, comprehensive, multimedia training manual for stem cell researchers. This manual has easy to follow step-by-step laboratory protocols and instructional videos as well as perspectives written by the world leaders in the field.
Editors: Gary S. Stein, Maria Borowski, Mai X. Luong, Meng-Jiao Shi, Kelly P. Smith, Priscilla Vazquez. 440 pages, Wiley-Blackwell. (January 2011)