Processing of equine bone marrow using the automated MarrowXpress System: RBC depletion, volume reduction, and mononuclear cell recovery.
Abstract: The therapeutic use of bone marrow-derived mononuclear cells (MNCs) and mesenchymal stem cells for the treatment of soft tissue and orthopedic injuries in equine patients is expanding. After collection, bone marrow must be reduced in volume and depleted of RBCs for immediate therapeutic use or to prepare cells for culture or cryopreservation and storage. The MarrowXpress (MXP) System is an automated, closed, sterile system designed to process human bone marrow samples. Objective: The purpose of this study was to evaluate the capacity of the MXP System to process equine bone marrow to reduce volume, deplete RBCs, and enhance recovery of MNCs. Methods: Bone marrow was collected from 47 horses into 2 60-mL syringes containing heparin and processed using the MXP System. HCT, total nucleated cell (TNC) count, and MNC count were obtained for each sample before and after processing using an Advia 120 hematology analyzer. Volume reduction, RBC depletion, and recovery of TNCs and MNCs were calculated. Results: For equine bone marrow samples, mean values were 73.2% for RBC depletion and 78.0% for volume reduction. TNC count before processing was 2.5 ± 1.2 × 10(7) and after processing was significantly higher at 7.8 ± 3.3 × 10(7) (P < .0001), with a recovery of 68.5 ± 24.5% (mean ± SD). MNC count before processing was 1.1 ± 0.9 × 10(7) and after processing was significantly higher at 3.8 ± 1.9 × 10(7) (P < .0001), with a recovery 73.0 ± 31.5%. Conclusions: The MXP System can reliably reduce volume and deplete RBCs from aspirates of equine bone marrow aspirates. MNCs can be recovered in a reproducible and sterile manner. Further studies evaluating the effects of the MXP System on cell viability, identification of mesenchymal stem cells (MSCs), and the efficacy of MSC expansion are warranted.
© 2011 American Society for Veterinary Clinical Pathology.
Publication Date: 2011-11-03 PubMed ID: 22092275DOI: 10.1111/j.1939-165X.2011.00368.xGoogle Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigated the application of the MarrowXpress (MXP) System, normally used for humans, in processing horse bone marrow in terms of reducing the overall volume, eliminating red blood cells (RBCs), and enhancing the recovery of mononuclear cells (MNCs).
Objective and Methodology
- The aim of the study was to assess the effectiveness of the MXP System in processing equine bone marrow. Specifically, the study looked at reducing the volume, depleting RBCs, and improving the recovery of MNCs.
- For this study, bone marrow was collected from 47 horses. The marrow was preserved in syringes containing heparin. After collection, the samples were processed using the MXP System.
- Scientists analyzed various factors pre and post processing. These included Hematocrit (HCT), Total nucleated cell (TNC) count, and MNC count. The volume reduction, RBC reduction, and recovery rate for TNC and MNC were all computed.
Results
- The study found that the MXP system was indeed effective in processing equine bone marrow samples. RBCs were depleted by an average of 73.2%. While there was an average volume reduction of 78.0%.
- The TNC count increased dramatically after processing. Before processing, the count was 2.5 ± 1.2 × 10(7) and after processing rose significantly to 7.8 ± 3.3 × 10(7). A recovery rate of 68.5 ± 24.5% was achieved.
- The MNC count also rose significantly after processing, from 1.1 ± 0.9 × 10(7) to 3.8 ± 1.9 × 10(7). The recovery stood at an impressive 73.0 ± 31.5%.
Conclusions
- The findings from the investigation establish that the MXP System can be successfully used to reduce the volume and deplete RBCs in samples of equine bone marrow aspirates.
- The system is also effective in recovering MNCs in a reproducible and sterile manner.
- However, as the researchers recommend further studies are needed to evaluate the effects of the MXP System on cell viability, the effect on mesenchymal stem cells (MSCs), and the efficacy of the MSC expansion process.
Cite This Article
APA
Owens SD, Burges J, Johns JL, Carrade DD, Galuppo LD, Librach F, Borjesson DL.
(2011).
Processing of equine bone marrow using the automated MarrowXpress System: RBC depletion, volume reduction, and mononuclear cell recovery.
Vet Clin Pathol, 40(4), 444-449.
https://doi.org/10.1111/j.1939-165X.2011.00368.x Publication
Researcher Affiliations
- Department of Pathology, Microbiology and Immunology.
- UC Davis Veterinary Blood Bank.
- Department of Pathology, Microbiology and Immunology.
- Department of Pathology, Microbiology and Immunology.
- Department of Surgical & Radiological Sciences, School of Veterinary Medicine, University of California-Davis, Davis, CA, USA.
- UC Davis Veterinary Blood Bank.
- Department of Pathology, Microbiology and Immunology.
MeSH Terms
- Animals
- Bone Marrow Purging / instrumentation
- Bone Marrow Purging / veterinary
- Cell Separation / instrumentation
- Cell Separation / veterinary
- Erythrocyte Count / veterinary
- Erythrocyte Volume / veterinary
- Erythrocytes / cytology
- Female
- Horses / blood
- Male
- Mesenchymal Stem Cells / cytology
- Monocytes / cytology
- Reproducibility of Results
Citations
This article has been cited 1 times.- Radtke CL, Nino-Fong R, Esparza Gonzalez BP, McD￮ LA. Application of a novel sorting system for equine mesenchymal stem cells (MSCs). Can J Vet Res 2014 Oct;78(4):290-6.
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