2008   Volume No 16 – pages 47-55

Title:Instant stem cell therapy: Characterization and concentration of human mesenchymal stem cells in vitro

Author: P Kasten, I Beyen, M Egermann, AJ Suda, AA Moghaddam, G Zimmermann, R Luginbühl

Address:Orthopaedic Surgery Hospital, University of Heidelberg, Schlierbacher Landstr. 200a, D-69118 Heidelberg, Germany

E-mail: Philip.Kasten@ok.uni-heidelberg.de

Key Words: Stem cell, bone regeneration, osteogenesis, tissue engineering, therapy, isolation protocol

Publication date: October 23rd 2008

Abstract: In regenerative medicine, there is an approach to avoid expansion of the mesenchymal stem cell (MSC) before implantation. The aim of this study was to compare methods for instant MSC therapy by use of a portable, automatic and closed system centrifuge that allows for the concentration of MSCs. The main outcome measures were the amount of MSCs per millilitre of bone marrow (BM), clusters of differentiation (CD), proliferation and differentiation capacities of the MSC. A volume reduction protocol was compared to the traditional laboratory methods of isolation using a Ficoll gradient and native BM. Fifty millilitres of BM were obtained from haematologically healthy male Caucasians (n=10, age 8 to 49 years). The number of colony forming units-fibroblast (CFU-F)/ml BM was highest in the centrifuge volume reduction protocol, followed by the native BM (not significant), the centrifuge Ficoll (p=0.042) and the manual Ficoll procedure (p=0.001). The MSC of all groups could differentiate into the mesenchymal lineages without significant differences between the groups. The CD pattern was identical for all groups: CD13+; CD 44+; CD73 +; CD90+; CD105+; HLA-A,B,C+; CD14-; CD34-; CD45-; CD271-; HLA-DR-. In a further clinical pilot study (n=5) with 297 ml BM (SD 18.6), the volume reduction protocol concentrated the MSC by a factor of 14: there were 1.08 x 102 MSC/ml BM (standard deviation (SD) 1.02 x 102) before concentration, 14.8 x 102 MSC/ ml BM (SD 12.4 x 102) after concentration, and on average 296 x 102 MSC (SD 248.9 x 102, range 86.4-691.5 x 102) were available for MSC therapy. The volume reduction protocol of the closed centrifuge allows for the highest concentration of the MSC, and therefore, is a promising candidate for instant stem cell therapy.

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