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. 2011 Mar;6(3):346-58.
doi: 10.1038/nprot.2010.199. Epub 2011 Feb 24.

Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells

Shigeki Sugii  1 , Yasuyuki Kida, W Travis Berggren, Ronald M Evans
Affiliations

Feeder-dependent and feeder-independent iPS cell derivation from human and mouse adipose stem cells

Shigeki Sugii et al. Nat Protoc. 2011 Mar.

Abstract

Adipose tissue is an abundantly available source of proliferative and multipotent mesenchymal stem cells with promising potential for regenerative therapeutics. We previously demonstrated that both human and mouse adipose-derived stem cells (ASCs) can be reprogrammed into induced pluripotent stem cells (iPSCs) with efficiencies higher than those that have been reported for other cell types. The ASC-derived iPSCs can be generated in a feeder-independent manner, representing a unique model to study reprogramming and an important step toward establishing a safe, clinical grade of cells for therapeutic use. In this study, we provide a detailed protocol for isolation, preparation and transformation of ASCs from fat tissue into mouse iPSCs in feeder-free conditions and human iPSCs using feeder-dependent or feeder/xenobiotic-free processes. This protocol also describes how ASCs can be used as feeder cells for maintenance of other pluripotent stem cells. ASC derivation is rapid and can be completed in <1 week, with mouse and human iPS reprogramming times averaging 1.5 and 2.5 weeks, respectively.

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Figures

Figure 1
Figure 1
Morphology of ADS cells in culture. (a) Mouse ADS cells grown in Expansion DMEM media. (b) Human ADS cells grown in hADSC media.
Figure 2
Figure 2
Derivation of mouse ADS-derived iPS cells. (a) An intermediate iPS colony at day 5 post-infection. Colonies are derived from mADS cells that were isolated from Oct4-EGFP mice. Note that the colony in the middle is weakly positive for GFP (right panel). (b) A developing iPS colony derived from Oct4-EGFP mADS cells at day 9. The colony is positive for GFP, indicating Oct4 activation. (c) An iPS colony (left) is picked and expanded in culture (right).
Figure 3
Figure 3
Derivation of human ADS-derived iPS cells. (a) Development of human iPS colonies at different stages. (b) Morphology of a hADS-derived iPS colony generated in the feeder-free condition. (c) A larger magnification of a hADS-derived iPS colony produced in the xeno-free condition. Note that xeno-free iPS cells generally appear flat.
See this image and copyright information in PMC

References

    1. Murry CE, Keller G. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell. 2008;132:661–680. - PubMed
    1. Muller R, Lengerke C. Patient-specific pluripotent stem cells: promises and challenges. Nat Rev Endocrinol. 2009;5:195–203. - PubMed
    1. Rodriguez-Piza I, et al. Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells under Xeno-free Conditions. Stem Cells. 2009;28:36–44. - PubMed
    1. Chidgey AP, Layton D, Trounson A, Boyd RL. Tolerance strategies for stem-cell-based therapies. Nature. 2008;453:330–337. - PubMed
    1. Martin MJ, Muotri A, Gage F, Varki A. Human embryonic stem cells express an immunogenic nonhuman sialic acid. Nat Med. 2005;11:228–232. - PubMed

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