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 ES Cells

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


Derivation of Induced Pluripotent Stem (iPS) Cells

TitleSourceMethod
Human iPS Cell Derivation/ReprogrammingDaley Lab, Children’s Hospital, BostonRetrovirus
Generation of Human induced Pluripotent Stem CellsCenter for iPS Cell Research and Application, Kyoto UniversityRetrovirus
Human iPS Cell Generation with hSTEMCCACenter for Regenerative Medicine, Boston UniversityLentivirus, STEMCCA
Human fibroblast reprogramming: Lentiviral method (hSTEMCCA) iPS Core Facility, Harvard UniversityLentivirus, STEMCCA
Reprogramming human fibroblasts using an episomal vector systemiPS Core Facility, Harvard UniversityEpisomal
Reprogramming Fibroblasts with SendaiMount Sinai School of MedicineSendai Virus
iPSC Reprogramming From Human Peripheral Blood Using Sendai Virus Mediated Gene TransferUniversity of PennsylvaniaSendai Virus

 

Basic Techniques for Growth of Pluripotent Stem Cells

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.

Culture Protocols from the NIH Stem Cell Unit
Details human ES cellculture on feeder and feeder-free as well as subcloning, EB differentiation, marker staining and FACS analysis.

Standard Operating Procedures (SOP) for the culture of human iPS
Culture protocols from the Harvard iPS Cell Core Facility

WiCell Protocols
Detailed set of PDFs with figures covering MEF feeder derivation and culture and the thawing, splitting, selection and maintenance of human ES cells.

Bresagen Protocols
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.

 

Other Resources

StemBook Protocols
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)

 

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