Extracellular Matrix-Dependent Generation of Integration- and Xeno-Free iPS Cells Using a Modified mRNA Transfection Method

Lee, Kang-In; Lee, Seo-Young; Hwang, Dong-Youn
January 2016
Stem Cells International;1/12/2016, p1
Academic Journal
Human induced pluripotent stem cells (iPS cells) hold great promise in the field of regenerative medicine, especially immune-compatible cell therapy. The most important safety-related issues that must be resolved before the clinical use of iPS cells include the generation of “footprint-free” and “xeno-free” iPS cells. In this study, we sought to examine whether an extracellular matrix- (ECM-) based xeno-free culture system that we recently established could be used together with a microRNA-enhanced mRNA reprogramming method for the generation of clinically safe iPS cells. The notable features of this method are the use of a xeno-free/feeder-free culture system for the generation and expansion of iPS cells rather than the conventional labor-intensive culture systems using human feeder cells or human feeder-conditioned medium and the enhancement of mRNA-mediated reprogramming via the delivery of microRNAs. Strikingly, we observed the early appearance of iPS cell colonies (~11 days), substantial reprogramming efficiency (~0.2–0.3%), and a high percentage of ESC-like colonies among the total colonies (~87.5%), indicating enhanced kinetics and reprogramming efficiency. Therefore, the combined method established in this study provides a valuable platform for the generation and expansion of clinically safe (i.e., integration- and xeno-free) iPS cells, facilitating immune-matched cell therapy in the near future.


Related Articles

  • A comparison of non-integrating reprogramming methods. Schlaeger, Thorsten M; Daheron, Laurence; Brickler, Thomas R; Entwisle, Samuel; Chan, Karrie; Cianci, Amelia; DeVine, Alexander; Ettenger, Andrew; Fitzgerald, Kelly; Godfrey, Michelle; Gupta, Dipti; McPherson, Jade; Malwadkar, Prerana; Gupta, Manav; Bell, Blair; Doi, Akiko; Jung, Namyoung; Li, Xin; Lynes, Maureen S; Brookes, Emily // Nature Biotechnology;Jan2015, Vol. 33 Issue 1, p58 

    Human induced pluripotent stem cells (hiPSCs) are useful in disease modeling and drug discovery, and they promise to provide a new generation of cell-based therapeutics. To date there has been no systematic evaluation of the most widely used techniques for generating integration-free hiPSCs....

  • Generation of iPSCs by Nonintegrative RNA-Based Reprogramming Techniques: Benefits of Self-Replicating RNA versus Synthetic mRNA. Steinle, Heidrun; Weber, Marbod; Behring, Andreas; Mau-Holzmann, Ulrike; Schlensak, Christian; Wendel, Hans Peter; Avci-Adali, Meltem // Stem Cells International;6/19/2019, p1 

    The reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is gaining in importance in the fields of regenerative medicine, tissue engineering, and disease modeling. Patient-specific iPSCs have as an unlimited cell source a tremendous potential for generating various types of...

  • Human iPS cell-derived alveolar epithelium repopulates lung extracellular matrix. Ghaedi, Mahboobe; Calle, Elizabeth A.; Mendez, Julio J.; Gard, Ashley L.; Balestrini, Jenna; Booth, Adam; Bove, Peter F.; Liqiong Gui; White, Eric S.; Niklason, Laura E. // Journal of Clinical Investigation;Nov2013, Vol. 123 Issue 11, p4950 

    The use of induced pluripotent stem cells (iPSCs) has been postulated to be the most effective strategy for developing patient-specific respiratory epithelial cells, which may be valuable for lung-related cell therapy and lung tissue engineering. We generated a relatively homogeneous population...

  • Hypothermic and cryogenic preservation of tissue‐engineered human bone. Tam, Edmund; McGrath, Madison; Sladkova, Martina; AlManaie, Athbah; Alostaad, Anaam; Peppo, Giuseppe Maria // Annals of the New York Academy of Sciences;Jan2020, Vol. 1460 Issue 1, p77 

    To foster translation and commercialization of tissue‐engineered products, preservation methods that do not significantly compromise tissue properties need to be designed and tested. Robust preservation methods will enable the distribution of tissues to third parties for research or...

  • Comparison of the Efficiency of Viral Transduction and Episomal Transfection in Human Fibroblast Reprogramming. Vdovin, A.; Lupatov, A.; Kholodenko, I.; Yarygin, K. // Bulletin of Experimental Biology & Medicine;Nov2015, Vol. 160 Issue 1, p123 

    Induced pluripotent cells were derived from adult human skin fibroblast by using two methods of reprogramming. Episomal transfection with vectors containing oriP/EBNA-1 sequence for delivery of reprogramming genes Oct4, Sox2, Klf4, L-Myc, and Lin28 proved to be more effective than viral...

  • THE CHANGING FACE OF THERAPEUTICS: AN INTRODUCTION TO STEM CELL AND REGENERATIVE THERAPIES. Seewoodhary, Jason // West London Medical Journal;Sep2011, Vol. 3 Issue 3, p28 

    Stem cell therapies are revolutionizing therapeutics by providing a curative approach to diseases for which, until very recently, symptomatic treatments were only available. This review will critically consider the multidisciplinary role, use and implications of embryonic, adult, and induced...

  • Tbx3: another important piece fitted into the pluripotent stem cell puzzle. Pirity, Melinda K.; Dinnyes, Andras // Stem Cell Research & Therapy;2010, Vol. 1 Issue 2, p1 

    Induced pluripotent stem cells (iPSCs) are novel tools for biomedical research, with a promise for future regenerative medicine applications. Recently, Han and colleagues reported in Nature that T box gene 3 (Tbx3) can improve the quality of mouse iPSCs and increase their germline transmission...

  • Patient-specific Induced Pluripotent Stem Cells as a Platform for Disease Modeling, Drug Discovery and Precision Personalized Medicine. Young, Wilson; D'Souza, Sunita L.; Lemischka, Ihor R.; Schaniel, Christoph // Journal of Stem Cell Research & Therapy;Special2012, Vol. 2 Issue S, Special section p1 

    The breakthrough development of induced pluripotent stem cell (iPSC) technology is not only revolutionizing basic stem cell science but is also spurring efforts to reprogram one somatic cell type directly into another. Induced pluripotent stem cells provide scientists with a self-renewing and,...

  • Advances and applications of induced pluripotent stem cells. Pietronave, Stefano; Prat, Maria // Canadian Journal of Physiology & Pharmacology;Mar2012, Vol. 90 Issue 3, p317 

    Direct reprogramming of somatic cells into pluripotent cells is an emerging technology for creating patient-specific cells, and potentially opens new scenarios in medical and pharmacological fields. From the discovery of Shinya Yamanaka, who first obtained pluripotent cells from fibroblasts by...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics