HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 2006-0015v1 24/10/2220    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints/Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Guo, K.-T. Articles by Wendel, H. P. Search for Related Content PubMed PubMed Citation Articles by Guo, K.-T. Articles by Wendel, H. P.

TECHNOLOGY DEVELOPMENT

A New Technique for the Isolation and Surface Immobilization of Mesenchymal Stem Cells from Whole Bone Marrow Using High-Specific DNA Aptamers

^aDepartment of Thoracic, Cardiac, and Vascular Surgery, University Hospital of Tuebingen;
^bInstitute of Clinical and Experimental Transfusion Medicine, University of Tuebingen, Tuebingen, Germany

Key Words. Mesenchymal stem cells • Aptamer • Fluorescence-activated cell sorting analysis

Correspondence: Hans P. Wendel, Ph.D., Calwer Str. 7/1, 72076 Tuebingen, Germany. Telephone: 07071/2986605; Fax: 07071/295369; e-mail: hp.wendel{at}uni-tuebingen.de

Received January 9, 2006; accepted for publication June 10, 2006.
First published online in STEM CELLS EXPRESS   June 22, 2006.



Adult mesenchymal stem cells (aMSCs) are a stem cell population present in bone marrow, which can be isolated and expanded in culture and characterized. Due to the lack of specific surface markers, it is difficult to separate the MSCs from bone marrow directly. Here, we present a novel method using high-specific nucleic acids called aptamers. Porcine MSCs were used as a target to generate aptamers by combinatorial chemistry out of a huge random library with in vitro technology called systematic evolution of ligands by exponential enrichment (SELEX). After cloning and sequencing, the binding affinity was detected using a cell-sorting assay with streptavidin-coated magnetic microbeads. We also used 12-well plates immobilized with aptamers to fish out MSCs from the cell solution and a fluorescein isothiocyanate-labeled aptamer to sort MSCs from bone marrow using high-speed fluorescence-activated cell sorting. The cells showed high potency to differentiate into osteogenic, as well as into adipogenic, lineages with typical morphological characteristics. Surface marker staining showed that the attached cells were CD29⁺, CD44⁺, CD45^–, CD90⁺, SLA class I⁺, SLA DQ^–, and SLA DR^–. Compared with existing methods, this study established a novel, rapid, and efficient method for direct isolation of aMSCs from porcine bone marrow by using an aptamer as a probe to fish out the aMSCs. This new application of aptamers can facilitate aMSC isolation and enrichment greatly, thereby enhancing the rate of aMSC-derived cells after in vitro differentiation for various applications in the emerging field of tissue engineering and regenerative medicine.