Author Disclosure Statement: The authors declare that no conflicting financial interests exist.
1552-4930/asset/olbannerleft.gif?v=1&s=cf6fcbfdc8ffa722551b71f8be0977b5b04db6ee)
1552-4930/asset/olbannerright.gif?v=1&s=6d0ba4e64946e5d9d4ebba4b8395a68bf324cd0b)
Original Article
You have free access to this content
Optimal reprogramming factor stoichiometry increases colony numbers and affects molecular characteristics of murine induced pluripotent stem cells†‡
Article first published online: 4 MAY 2011
DOI: 10.1002/cyto.a.21072
Copyright © 2011 International Society for Advancement of Cytometry
Additional Information
How to Cite
Tiemann, U., Sgodda, M., Warlich, E., Ballmaier, M., Schöler, H. R., Schambach, A. and Cantz, T. (2011), Optimal reprogramming factor stoichiometry increases colony numbers and affects molecular characteristics of murine induced pluripotent stem cells. Cytometry, 79A: 426–435. doi: 10.1002/cyto.a.21072
- †
- ‡
Author contributions: U.T.: vector production, iPSC generation and characterization, data analysis and interpretation, and manuscript writing. M.S.: qRT-PCR analyses. E.W.: generation of vector plasmids. M.B.: flow cytometry setup and cell sorting. H.R.S.: conception and design. A.S.: generation of vector plasmids and design of the study. T.C.: conceptional design of the study, data analysis and interpretation, and manuscript writing. All authors approved the final version of the manuscript.
Publication History
- Issue published online: 18 MAY 2011
- Article first published online: 4 MAY 2011
- Manuscript Accepted: 4 APR 2011
- Manuscript Revised: 25 MAR 2011
- Manuscript Received: 2 FEB 2011
Funded by
- German Research Foundation. Grant Number: DFG: EXC 62/1
- German Ministry for Education and Research. Grant Number: 01GN0812, 01GM0854 and ReGene
- DAAD. Grant Number: 0315187
LITERATURE CITED
- 1,. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006; 126: 663–676.
- 2,,,,,,,. In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature 2007; 448: 318–324.
- 3,,,. c-Myc is dispensable for direct reprogramming of mouse fibroblasts. Cell Stem Cell 2008; 2: 10–12.
- 4,,,,,,,. Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol 2008; 26: 1269–1275.
- 5,,,,,,,,,, , , , , , , , . Oct4-induced pluripotency in adult neural stem cells. Cell 2009; 136: 411–419.
- 6,,,,,,,,,. 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; 106: 12759–12764.
- 7,,. A guide to choosing fluorescent proteins. Nat Methods 2005; 2: 905–909.
- 8,,,,,,,,,, , , . Lentiviral vector design and imaging approaches to visualize the early stages of cellular reprogramming. Mol Ther 2011; 19: 782–789.
- 9,,,,,,. Equal potency of gammaretroviral and lentiviral SIN vectors for expression of O6-methylguanine-DNA methyltransferase in hematopoietic cells. Mol Ther 2006; 13: 391–400.
- 10,,,,,,,,. Aberrant silencing of imprinted genes on chromosome 12qF1 in mouse induced pluripotent stem cells. Nature 2010; 465: 175–181.
- 11,. Emerging use of stem cells in regenerative medicine. Biochem J 2010; 428: 11–23.
- 12,. Induced pluripotent stem cells: Characteristics and perspectives. Adv Biochem Eng Biotechnol 2010; 123: 107–126.
- 13,,,,,,,,,. Disease-specific induced pluripotent stem cells. Cell 2008; 134: 877–886.
- 14,,,,,,. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007; 131: 861–872.
- 15,,,,,,,,,, , . Induced pluripotent stem cell lines derived from human somatic cells. Science 2007; 318: 1917–1920.
- 16,,,,. Generation of mouse induced pluripotent stem cells without viral vectors. Science 2008; 322: 949–953.
- 17,,,,. Induced pluripotent stem cells generated without viral integration. Science 2008; 322: 945–949.
- 18,,,,,,. Human induced pluripotent stem cells free of vector and transgene sequences. Science 2009; 324: 797–801.
- 19,. Adenoviral gene delivery can reprogram human fibroblasts to induced pluripotent stem cells. Stem Cells 2009; 27: 2667–2674.Direct Link:
- 20,,,,,. Dual-color flow cytometric detection of fluorescent proteins using single-laser (488-nm) excitation. Cytometry 1998; 31: 147–152.Direct Link:
- 21,,. Simultaneous flow cytometric analyses of enhanced green and yellow fluorescent proteins and cell surface antigens in doubly transduced immature hematopoietic cell populations. Cytometry 2000; 40: 126–134.Direct Link:
- 22,,. Analysis of violet-excited fluorochromes by flow cytometry using a violet laser diode. Cytometry Part A 2003; 54A: 48–55.Direct Link:
- 23,. Fluorescent protein tools for studying protein dynamics in living cells: A review. J Biomed Opt 2008; 13: 031202.
- 24,,,. Dynamic single-cell imaging of direct reprogramming reveals an early specifying event. Nat Biotechnol 2010; 28: 521–526.
- 25,,,,,,. Direct reprogramming of human neural stem cells by OCT4. Nature 2009; 461: 648–653.
- 26,,,,,,,. Role of the murine reprogramming factors in the induction of pluripotency. Cell 2009; 136: 364–377.
- 27,,,,,. Transcriptional signature and memory retention of human-induced pluripotent stem cells. PLoS One 2009; 4: e7076.
- 28,,,,,,,,,. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol 2007; 26: 101–106.
- 29,,,,. Efficient induction of transgene-free human pluripotent stem cells using a vector based on Sendai virus, an RNA virus that does not integrate into the host genome. Proc Jpn Acad Ser B Phys Biol Sci 2009; 85: 348–362.
- 30,,,,,,,,,, , . A nonviral minicircle vector for deriving human iPS cells. Nat Methods 2010; 7: 197–199.
- 31,,,,,,,,,, , , , . Generation of induced pluripotent stem cells using recombinant proteins. Cell Stem Cell 2009; 4: 381–384.
- 32,,,,,,,,,, . Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell Stem Cell 2009; 4: 472–476.
- 33,,,,,,,,,, , , , , , . Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 2010; 7: 618–630.