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Open Access Research article

Isolation of equine multipotent mesenchymal stromal cells by enzymatic tissue digestion or explant technique: comparison of cellular properties

Claudia Gittel1*, Walter Brehm1, Janina Burk1, Henriette Juelke2, Carsten Staszyk3 and Iris Ribitsch14

Author Affiliations

1 Large Animal Clinic for Surgery, University of Leipzig, An den Tierkliniken 21, 04103 Leipzig, Germany

2 Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Philipp-Rosenthal-Straße 55, 04103 Leipzig, Germany

3 Department of Veterinary-Anatomy, -Histology and -Embryology, Faculty for Veterinary Medicine, Justus-Liebig-University Giessen, Frankfurter Str. 98, 35392 Giessen, Germany

4 Equine Hospital, Clinic for Equine Surgery, University of Veterinary Medicine, Veterinärplatz 1, 1210 Vienna, Austria

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BMC Veterinary Research 2013, 9:221  doi:10.1186/1746-6148-9-221

Published: 29 October 2013

Abstract

Background

The treatment of tendon lesions with multipotent mesenchymal stromal cells (MSCs) is widely used in equine medicine. Cell sources of MSCs include bone marrow, as well as solid tissues such as adipose tissue. MSCs can be isolated from these solid tissues either by enzymatic digestion or by explant technique. However, the different preparation techniques may potentially influence the properties of the isolated MSCs. Therefore, the aim of this study was to investigate and compare the effects of these two different methods used to isolate MSCs from solid tissues.

Equine adipose tissue, tendon and umbilical cord matrix served as solid tissue sources of MSCs with different stiffness and density. Subsequent to tissue harvest, MSCs were isolated either by enzymatic digestion with collagenase or by explant technique. Cell yield, growth, differentiation potential and tendon marker expression were analysed.

Results

At first passage, the MSC yield was significantly higher in enzymatically digested tissue samples than in explanted tissue samples, despite a shorter period of time in primary culture. Further analysis of cell proliferation, migration and differentiation revealed no significant differences between MSCs isolated by enzymatic digestion and MSCs isolated by explant technique. Interestingly, analysis of gene expression of tendon markers revealed a significantly higher expression level of scleraxis in MSCs isolated by enzymatic digestion.

Conclusions

Both isolation techniques are feasible methods for successful isolation of MSCs from solid tissues, with no major effects on cellular proliferation, migration or differentiation characteristics. However, higher MSC yields were achieved in a shorter period of time by collagenase digestion, which is advantageous for the therapeutic use of MSCs. Moreover, based on the higher level of expression of scleraxis in MSCs isolated by enzymatic digestion, these cells might be a better choice when attempting tendon regeneration.

Keywords:
Horse; Regenerative medicine; Collagenase; Cell isolation; Scleraxis