close
Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Sep 17;285(38):29270-8.
doi: 10.1074/jbc.M109.055889. Epub 2010 Jul 1.

Induction of pluripotent stem cells from human third molar mesenchymal stromal cells

Yasuaki Oda  1 , Yasuhide Yoshimura, Hiroe Ohnishi, Mika Tadokoro, Yoshihiro Katsube, Mari Sasao, Yoko Kubo, Koji Hattori, Shigeru Saito, Katsuhisa Horimoto, Shunsuke Yuba, Hajime Ohgushi
Affiliations

Induction of pluripotent stem cells from human third molar mesenchymal stromal cells

Yasuaki Oda et al. J Biol Chem. .

Abstract

The expression of four transcription factors (OCT3/4, SOX2, KLF4, and MYC) can reprogram mouse as well as human somatic cells to induced pluripotent stem (iPS) cells. We generated iPS cells from mesenchymal stromal cells (MSCs) derived from human third molars (wisdom teeth) by retroviral transduction of OCT3/4, SOX2, and KLF4 without MYC, which is considered as oncogene. Interestingly, some of the clonally expanded MSCs could be used for iPS cell generation with 30-100-fold higher efficiency when compared with that of other clonally expanded MSCs and human dermal fibroblasts. Global gene expression profiles demonstrated some up-regulated genes regarding DNA repair/histone conformational change in the efficient clones, suggesting that the processes of chromatin remodeling have important roles in the cascade of iPS cells generation. The generated iPS cells resembled human embryonic stem (ES) cells in many aspects, including morphology, ES marker expression, global gene expression, epigenetic states, and the ability to differentiate into the three germ layers in vitro and in vivo. Because human third molars are discarded as clinical waste, our data indicate that clonally expanded MSCs derived from human third molars are a valuable cell source for the generation of iPS cells.

PubMed Disclaimer

Figures

FIGURE 1.
FIGURE 1.
Generation of iPS cells from MSCs. A, reprogramming efficiency of non-cloned parental MSCs from the third molars of 10-, 16-, and 13-year-old donors. The bars represent the S.D. (n = 10). B, proliferation rate of clonally expanded MSCs from 10-year-old donor and HDF at day 5. The bars indicate S.D. (n = 3). C, Reprogramming efficiency of clonally expanded MSCs and HDF cells. The closed circles represent average of two independent experiments, and the bars indicate S.D. (n = 8). D, ALP staining of three factors transduced cells at day 30. E, viral transduction efficiency of the pMXs retroviral vector. The percentages indicate the ratio of DsRed-Express-positive cells. F, parental cells of clonally expanded MSC and HDF cells (upper panel), iPS cells from the parental cells (middle panel), and ALP staining of each iPS cell (lower panel). Scale bars = 100 μm.
FIGURE 2.
FIGURE 2.
Characterization of iPS cells from clonally expanded MSCs and HDF. A, immunocytochemistry of SSEA-3, SSEA-4, TRA-1–60, TRA-1–81, OCT3/4, and NANOG for iPS cells. Scale bars = 100 μm. B, DNA methylation states of the OCT3/4 and NANOG promoter regions. Open circles indicate unmethylated CpGs, and closed circles indicate methylated CpGs. C, RT-PCR analysis of ES cell marker genes and retroviral Tgs for iPS cells from different colonies (cl.58/cl.15 derived from 10F-15 clonal cells, cl.5/cl.8 derived from 10F-101 clonal cells, and cl.1/cl.8 derived from HDF). D, telomerase activity detected by TRAP assay. +, heat-treated; −, non-treated samples; P, parental cell. IC, internal control.
FIGURE 3.
FIGURE 3.
In vitro differentiation of iPS cells from clonally expanded MSCs and HDF. A, EB formation of iPS cells at day 8. B–E, immunocytochemistry for SOX17 (B), AFP (green) (C), desmin (red) (D), and α-smooth muscle actin βIII-tubulin (E). F, immunocytochemistry of differentiated iPS cells cultured on PA6 feeder cells at day 18. Tyrosine hydroxylase- and βIII-tubulin-positive cells are indicated as green and red cells, respectively. Nuclei were stained with Hoechst 33342 (blue). Scale bars = 100 μm. G, RT-PCR analysis of three germ layer marker genes. P, parental cell; U, undifferentiated iPS cells; D, differentiated iPS cells (EB).
FIGURE 4.
FIGURE 4.
Teratoma formation after in vivo implantation of iPS cells from clonally expanded MSCs and HDF. A–I, hematoxylin and eosin staining of harvested tumors. Each tumor (teratoma) contained three embryonic germ layer tissues including gut-like epithelium (A–C: endoderm), cartilage (D–F: mesoderm), and neuroepithelial rosettes (G–I: ectoderm). cl.58, colony derived from 10F-15 clonal cells; cl.8, colony derived from 10F-101 clonal cells; cl.1, colony derived from HDF.
FIGURE 5.
FIGURE 5.
Global gene expression analysis of iPS cells from clonally expanded MSCs and their parental cells. A, expression levels of reprogramming genes in parental cells and iPS cells. cl.39, colony derived from 10F-15 clonal cells; cl.58, colony derived from 10F-15 clonal cells; cl.5, colony derived from 10F-101 clonal cells; cl.8, colony derived from 10F-101 clonal cells; cl.1, colony derived from HDF. B, global gene expression comparison between parental cell and iPS cell. Red circles indicate the expression levels of OCT3/4, NANOG, SALL4, LIN28, and TDGF1. Red lines indicate the equivalent and 3-fold differences between the two samples. C, comparison of gene expression profiles of parental cells, iPS cells, and human ES (hES) cells. cl.8, colony derived from HDF.
See this image and copyright information in PMC

References

    1. Takahashi K., Yamanaka S. (2006) Cell 126, 663–676 - PubMed
    1. Takahashi K., Tanabe K., Ohnuki M., Narita M., Ichisaka T., Tomoda K., Yamanaka S. (2007) Cell 131, 861–872 - PubMed
    1. Yu J., Vodyanik M. A., Smuga-Otto K., Antosiewicz-Bourget J., Frane J. L., Tian S., Nie J., Jonsdottir G. A., Ruotti V., Stewart R., Slukvin I. I., Thomson J. A. (2007) Science 318, 1917–1920 - PubMed
    1. Park I. H., Zhao R., West J. A., Yabuuchi A., Huo H., Ince T. A., Lerou P. H., Lensch M. W., Daley G. Q. (2008) Nature 451, 141–146 - PubMed
    1. Lowry W. E., Richter L., Yachechko R., Pyle A. D., Tchieu J., Sridharan R., Clark A. T., Plath K. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 2883–2888 - PMC - PubMed

Publication types

Substances