Equine bone marrow volume reduction, red blood cell depletion, and mononuclear cell recovery using the PrepaCyte-CB processing system.
Abstract: Volume reduction and RBC depletion of equine bone marrow specimens are necessary processing steps for the immediate therapeutic use of bone marrow (BM)-derived mesenchymal stem cells (MSC), and for MSC expansion in culture. Objective: The purpose of the study was to evaluate the ability of the PrepaCyte-CB processing system to reduce volume, deplete RBC, and recover mononuclear cells (MNC) from equine BM specimens. Methods: One hundred and twenty mL of heparinized BM were obtained from each of 90 horses. A CBC was performed on the BM pre- and post-PrepaCyte-CB processing. Volume and RBC reduction, and total nucleated cell (TNC) and MNC recoveries were determined. Results: Bone marrow volume was reduced from 120 mL to 21 mL with a median RBC depletion of 90.1% (range, 62.0-96.7%). The median preprocessing total TNC count was 2.2 × 10(9) (range, 0.46-7.9 × 10(9)) and the median postprocessing TNC count was 1.7 × 10(9) (range, 0.3-4.4 × 10(9); P < .0001), with a median recovery of 73.5% (range, 22.4-216.7%). The median preprocessing total MNC count was 0.9 × 10(9) (range, 0.1-4.7 × 10(9)) and median postprocessing total MNC count was 0.8 × 10(9) (range, 0.1-2.7 × 10(9); P = .06), with a median recovery of 83.7% (range, 15.4-413.9%). Conclusions: The PrepaCyte-CB processing system can be used to deplete both volume and RBC, and recover MNC from equine BM specimens. Further studies assessing the viability of MSC and the efficacy of MSC expansion after using the PrepaCyte-CB processing system are warranted.
© 2015 American Society for Veterinary Clinical Pathology.
Publication Date: 2015-01-27 PubMed ID: 25627833DOI: 10.1111/vcp.12236Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Biotechnology
- Bone Marrow
- Cell Proliferation
- Cell Viability
- Clinical Pathology
- Clinical Study
- Diagnosis
- Disease Diagnosis
- Disease Treatment
- Equine Health
- Equine Science
- Hematology
- Heparin
- In Vitro Research
- Laboratory Methods
- Mesenchymal Cells
- Mononuclear Cells
- Red Blood Cells
- Stem Cells
- Veterinary Medicine
- Veterinary Research
Summary
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This study investigates the effectiveness of the PrepaCyte-CB processing system in reducing the volume and red blood cell content of horse bone marrow samples while ensuring the recovery of mononuclear cells necessary for therapeutic use or culture expansion of mesenchymal stem cells.
Objective
- The research aims to analyze the PrepaCyte-CB processing system’s capability to decrease bone marrow volume, lessen the number of red blood cells, and ensure the retrieval of mononuclear cells from horse bone marrow samples. These processes are particularly crucial for therapeutic applications and expansion of bone marrow-derived mesenchymal stem cells in the lab.
Methodology
- A total of 120 mL of heparinized bone marrow was collected from 90 horses and subjected to a complete blood count both before and after the PrepaCyte-CB processing phase. This study sets out to monitor changes in bone marrow volume, red blood cell depletion, total nucleated cell (TNC), and mononuclear cell (MNC) recoveries.
Results
- The results showed that the bone marrow volume was successfully decreased from 120 mL to 21 mL, with a median red blood cell depletion reaching up to 90.1%. The study also noted substantial recoveries for both TNC and MNC. The median total TNC recovery was calculated at 73.5% decrease, while the median total MNC count observed an 83.7% recovery rate.
Conclusions
- The researchers conclude the effectiveness of the PrepaCyte-CB processing system in successfully reducing both bone marrow volume and red blood cell content and in ensuring the successful recovery of mononuclear cells from horse bone marrow samples. This may have potential implications for the therapeutic use of mesenchymal stem cells, however, to confirm this, additional research is recommended into the viability and efficacy of mesenchymal stem cell expansion following the use of the PrepaCyte-CB processing system.
Cite This Article
APA
Burton AG, Clark KC, Borjesson DL, Carrade DD, Burges J, Owens SD.
(2015).
Equine bone marrow volume reduction, red blood cell depletion, and mononuclear cell recovery using the PrepaCyte-CB processing system.
Vet Clin Pathol, 44(2), 188-193.
https://doi.org/10.1111/vcp.12236 Publication
Researcher Affiliations
- Veterinary Medical Teaching Hospital, University of California-Davis, Davis, CA, USA.
- Regenerative Medicine Laboratory, University of California-Davis, Davis, CA, USA.
- Department of Pathology, Microbiology and Immunology, University of California-Davis, Davis, CA, USA.
- Department of Pathology, Microbiology and Immunology, University of California-Davis, Davis, CA, USA.
- Regenerative Medicine Laboratory, University of California-Davis, Davis, CA, USA.
- Department of Pathology, Microbiology and Immunology, University of California-Davis, Davis, CA, USA.
MeSH Terms
- Animals
- Bone Marrow / chemistry
- Bone Marrow Purging / instrumentation
- Bone Marrow Purging / veterinary
- Cell Separation / instrumentation
- Cell Separation / veterinary
- Erythrocyte Count / veterinary
- Erythrocyte Volume / veterinary
- Erythrocytes / cytology
- Erythrocytes / physiology
- Horses / physiology
- Leukocytes, Mononuclear / cytology
- Leukocytes, Mononuclear / physiology
- Specimen Handling
Citations
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