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Open Access Research article

Murine mesenchymal progenitor cells from different tissues differentiated via mesenchymal microspheres into the mesodermal direction

Florian Böhrnsen13, Ulrich Lindner3, Markus Meier3, Abdelalim Gadallah1, Peter Schlenke2, Hendrik Lehnert3, Jürgen Rohwedel1 and Jan Kramer13*

Author affiliations

1 Institute of Virology and Cell Biology, University of Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany

2 Institute for Transfusion Medicine and Transplantation Immunology, University of Münster, Domagkstraße 11, 48149 Münster, Germany

3 Medical Department I, University of Lübeck, Ratzeburger Allee 160, 23538 Lübeck, Germany

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Citation and License

BMC Cell Biology 2009, 10:92  doi:10.1186/1471-2121-10-92

Published: 19 December 2009

Abstract

Background

Because specific marker molecules for phenotypical identification of mesenchymal stem and progenitor cells are missing, the assessment of the in vitro-differentiation capacity is a prerequisite to characterize these cells. However, classical differentiation protocols are often cell-consuming and time intensive. Therefore, the establishment of novel strategies for differentiation is one topic of current efforts in stem cell biology. The goal of this study was to demonstrate the practicability of a new differentiation test using plastic adherent cell isolates from different tissues.

Results

We introduced the mesenchymal microsphere method as a feasible time- and cell saving screening method to analyse multilineage differentiation properties of adult progenitor cells in a three-dimensional system. For this purpose we isolated, characterized and analyzed new sources of adult murine mesenchymal progenitor cells from perirenal adipose tissue and mediastinal stromal tissue in comparison to bone marrow progenitor cells. The proliferation capacity of the cells was demonstrated by determination of the daily doubling index. Although the flow cytometry analysis of undifferentiated cells revealed differences in the expression of CD marker molecules, all isolates have the capacity for multilineage differentiation following the mesenchymal microsphere protocol as well as the classical "micro mass body" protocol for chondrogenic and the monolayer cultivation protocol for osteogenic and adipogenic differentiation. Differentiation was characterized using histochemical and immunhistochemical staining as well as RT-PCR.

Conclusions

We were able to show that the mesenchymal microsphere method is an efficient test system for chondro-, osteo- and adipogenic differentiation of adult progenitor cells. The advantage of this system in comparison to classical protocols is that approximately 7 times lower cell numbers are necessary. Since classical culture procedures are time intensive because high cell numbers have to be obtained, the new differentiation method may also save cells and time in future clinical applications using human mesenchymal stromal cells.