Welcome!
The SKI Stem Cell Research Facility was created to expand and characterize human embryonic stem cells (hESCs) derived in the Tri-Institutional Community. We also produce iPS cells on demand on a fee-for-service basis. In addition to transferring pluirpotent stem cells, we provide:
• Training in hESC culture
• Transgenic hESCs and services
• Differentiated cell types derived from human or mouse ESCs
New technologies such as BAC transgenesis and homologous recombination are being refined for use in human pluripotent stem cells (Placantonakis et al. 2009; Brunet et al. 2009). The facility is also optimizing hESC culture in an effort to discover conditions more suitable for long-term maintenance and karyotypic stability of ESCs (Joubin et al. 2012).
Inventory
A number of human ESC lines from WiCell (H1, H7, H9, H13.B, H14, WA25, WA26 and WA27), two NIH registry lines from the Brivanlou Lab (Rockefeller University: RU01 and RU02) and ESI-017 and ESI-035 from BioTime, Inc. are available for transfer after the appropriate MTAs have been signed by the IP holder.
A number of human iPSC lines created in our facility are also available for distribution.
Cell Types
The facility derives neural, mesenchymal, endoderm, mesoderm, and trophectoderm from human pluripotent stem cells. For some cell types, we can also use mouse pluripotent stem cells. Contact us to discuss the production of specific cell types.
iPS
Our lab initially collaborated with the Sadelain and Studer labs (Papapetrou et al., 2009) to learn how to reprogram somatic cells the production of iPSCs. We typically use the Sendai viral vector method now to create iPSCs although we still offer lentiviral constructs for those that want that system.
Genetic Modification
In collaboration with Lorenz Studer’s lab, we were the first to make BAC transgenic reporter human embryonic stem cells (Placantonakis et al., 2009: see publications). We continue to develop BAC technologies in the lab, hoping to reduce the amount of time and money required to engineer successful lines. In addition to BACs, we routinely produce stable cell lines with conventional plasmids for constitutive expression or to knockdown your gene of interest with shRNAs.
Training
Human embryonic stem cell culture is challenging. We offer a three day, hands-on lab course that teaches basic hESC culture techniques. Topics covered include: hES thawing, expansion, passaging, and freezing. Participants will learn to maintain undifferentiated cells and to remove contaminating differentiated cells.
Protocols
Click links to view protocols. To download, ctrl-click [Mac] or right click [Windows] on link.
Maintenance of human ESCs
Endoderm
Mesoderm
Neural induction-preparing MS-5
BAC retrofitting and introduction into human and mouse ESCs
Publication
Kushwaha R, Jagadish N, Kustagi M, Tomishima MJ, Mendiratta G, Bansal M, Kim HR, Sumazin P, Alvarez MJ, Lefebvre C, Villagrasa-Gonzalez P, Viale A, Korkola JE, Houldsworth J, Feldman DR, Bosl GJ, Califano A, Chaganti RS.Interrogation of a context-specific transcription factor network identifies novel regulators of pluripotency.Stem Cells. 2015 Feb;33(2):367-77.
Kushwaha R, Thodima V, Tomishima MJ, Bosl GJ, Chaganti RS. miR-18b and miR-518b Target FOXN1 during epithelial lineage differentiation in pluripotent cells.Stem Cells Dev. 2014 May 15;23(10):1149-56.
Miller JD, Ganat YM, Kishinevsky S, Bowman RL, Liu B, Tu EY, Mandal PK, Vera E, Shim JW, Kriks S, Taldone T, Fusaki N, Tomishima MJ, Krainc D, Milner TA, Rossi DJ, Studer L. Human iPSC-based modeling of late-onset disease via progerin-induced aging. Cell Stem Cell. 2013 Dec 5;13(6):691-705.
Chambers SM, Mica Y, Lee G, Studer L, Tomishima MJ. Dual-SMAD Inhibition/WNT Activation-Based Methods to Induce Neural Crest and Derivatives from Human Pluripotent Stem Cells. Methods Mol Biol. 2013 Dec 4.
Gerhardt J, Tomishima MJ, Zaninovic N, Colak D, Yan Z, Zhan Q, Rosenwaks Z, Jaffrey SR, Schildkraut CL. The DNA replication program is altered at the FMR1 locus in fragile X embryonic stem cells. Mol Cell. 2014 Jan 9;53(1):19-31.
Harel S, Tu EY, Weisberg S, Esquilin M, Chambers SM, Liu B, Carson CT, Studer L, Reizis B, Tomishima MJ. ZFX controls the self-renewal of human embryonic stem cells. PLoS One. 2012;7(8):e42302.
Mica Y, Lee G, Chambers SM, Tomishima MJ, Studer L. Modeling neural crest induction, melanocyte specification, and disease-related pigmentation defects in hESCs and patient-specific iPSCs.Cell Rep. 2013 Apr 25;3(4):1140-52.
Piganeau M, Ghezraoui H, De Cian A, Guittat L, Tomishima M, Perrouault L, René O, Katibah GE, Zhang L, Holmes MC, Doyon Y, Concordet JP, Giovannangeli C, Jasin M, Brunet E. Cancer translocations in human cells induced by zinc finger and TALE nucleases. Genome Res. 2013 Jul;23(7):1182-93.
Ganat YM, Calder EL, Kriks S, Nelander J, Tu EY, Jia F, Battista D, Harrison N, Parmar M, Tomishima MJ, Rutishauser U, Studer L. Identification of embryonic stem cell-derived midbrain dopaminergic neurons for engraftment. J Clin Invest. 2012 Aug;122(8):2928-39.
Joubin K, Richardson A, Novoa N, Tu E, Tomishima MJ. The endothelial cell line bEnd.3 maintains human pluripotent stem cells. Stem Cells Dev. 2012 Aug 10;21(12):2312-21.
Chambers SM, Mica Y, Studer L, Tomishima MJ. Converting human pluripotent stem cells to neural tissue and neurons to model neurodegeneration. Methods Mol Biol. 2011;793:87-97.
Hoya-Arias R, Tomishima M, Perna F, Voza F, Nimer SD. L3MBTL1 deficiency directs the differentiation of human embryonic stem cells toward trophectoderm. Stem Cells Dev. 2011 Nov;20(11):1889-900.
Fasano CA, Chambers SM, Lee G, Tomishima MJ, Studer L. Efficient derivation of functional floor plate tissue from human embryonic stem cells. Cell Stem Cell. 2010 Apr 2;6(4):336-47.
Lee G, Chambers SM, Tomishima MJ, Studer L. Derivation of neural crest cells from human pluripotent stem cells. Nat Protoc. 2010 Apr;5(4):688-701. Epub 2010 Mar 18.
James D, Nam HS, Seandel M, Nolan D, Janovitz T, Tomishima M, Studer L, Lee G, Lyden D, Benezra R, Zaninovic N, Rosenwaks Z, Rabbany SY, Rafii S. Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFbeta inhibition is Id1 dependent. Nat Biotechnol. 2010 Jan 17. [
Lee G, Papapetrou EP, Kim H, Chambers SM, Tomishima MJ, Fasano CA, Ganat YM, Menon J, Shimizu F, Viale A, Tabar V, Sadelain M, Studer L. Modelling pathogenesis and treatment of familial dysautonomia using patient-specific iPSCs. Nature. 2009 Sep 17;461(7262):402-6.
Brunet E, Simsek D, Tomishima M, DeKelver R, Choi VM, Gregory P, Urnov F, Weinstock DM, Jasin M. Chromosomal translocations induced at specified loci in human stem cells. Proc Natl Acad Sci U S A. 2009 Jun 30;106(26):10620-5.
Papapetrou EP, Tomishima MJ, Chambers SM, Mica Y, Reed E, Menon J, Tabar V, Mo Q, Studer L, Sadelain M. Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation. Proc Natl Acad Sci U S A. 2009 Jun 23.
Chambers SM, Fasano CA, Papapetrou EP, Tomishima M, Sadelain M, Studer L. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009 Mar;27(3):275-80.
Placantonakis DG*, Tomishima MJ*, Lafaille F**, Desbordes SC**, Jia F, Socci ND, Viale A, Lee H, Harrison N, Tabar V, Studer L. BAC transgenesis in human ES cells as a novel tool to define the human neural lineage. Stem Cells. 2009 Mar;27(3):521-32.
Tabar V, Tomishima M, Panagiotakos G, Wakayama S, Menon J, Chan B, Mizutani E, Al-Shamy G, Ohta H, Wakayama T, Studer L. Therapeutic cloning in individual parkinsonian mice. Nat Med. 2008 Apr;14(4):379-81.
Bradbury MS, Panagiotakos G, Chan BK, Tomishima M, Zanzonico P, Vider J, Ponomarev V, Studer L, Tabar V. Optical bioluminescence imaging of human ES cell progeny in the rodent CNS. J Neurochem. 2007 Sep;102(6):2029-39.
Tomishima MJ, Hadjantonakis AK, Gong S, Studer L. Production of green fluorescent protein transgenic embryonic stem cells using the GENSAT bacterial artificial chromosome library. Stem Cells. 2007 Jan;25(1):39-45.
Book chapters
Placantonakis DG, Tomishima MJ, Lafaille FG, Studer L. 2008. Genetic manipulation of human embryonic stem cells. In Regulatory Networks in Stem Cells (eds. R. Vinagolu & V. Mohan). Humana Press, Totowa, NJ.
Tomishima MJ, Studer L. 2008. Embryonic stem cell-based models of Parkinson’s disease. In Parkinson’s Disease: Molecular and Therapeutic Insights from Model Systems (eds. R. Nass & S. Przedborski). Academic Press, San Diego, CA.
Tomishima MJ, Studer L. 2003. Embryonic stem cell differentiation into neural cells. In Neural Stem Cells: Development and Transplantation (ed. J.E. Bottenstein). Kluwer Academic Publishers, Boston, MA.
Dr. Ting Zhou, Ph.D. | Facility Manager
212.639.3913 [office]
212.639.3969 [lab]
zhout@mskcc.org
| Hours | Location |
|
9:30AM to 4:30PM Monday through Friday |
1275 York Avenue New York, New York 10065 |
