Performance of a gravitational marrow separator, multidirectional bone marrow aspiration needle, and repeated bone marrow collections on the production of concentrated bone marrow and separation of mesenchymal stem cells in horses.
Abstract: Objective-To determine the efficiency of a novel point-of-care gravitational marrow separator and bone marrow aspiration needle for concentrated bone marrow production and bone marrow-derived mesenchymal stem cell (MSC) separation and assess the effect of repeated bone marrow collections in horses. Animals-8 healthy adult horses. Procedures-Bone marrow aspiration was performed twice (1 month apart) from sternebral bodies with a standard or prototype multidirectional needle. Concentrated bone marrow was obtained by gravitational marrow separation and evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, MSCs, and cell viability. Results-Concentrated bone marrow samples obtained with the marrow separator had 5- to 19-fold bone marrow-derived MSC, WBC, and platelet counts, compared with original bone marrow samples. Use of a multidirectional needle increased the frequency of obtaining MSC-richer concentrated bone marrow. Repeating bone marrow aspiration at 1 month yielded greater MSC numbers but slightly lower cell viability after processing. Conclusions and Clinical Relevance-The gravitational bone marrow separator and multidirectional needle were used to effectively harvest bone marrow and improve the quality of concentrated bone marrow. Comparable, or even greater, numbers of bone marrow-derived MSCs were collected by repeated bone marrow aspiration after a 1-month interval from the same aspiration sites. Use of the marrow separator and multidirectional bone marrow aspiration needle can facilitate a 1-step, point-of-care, nonlaboratory method to obtain concentrated bone marrow as a mixture of bone marrow-derived MSCs and growth factors from platelets and plasma.
Publication Date: 2013-05-31 PubMed ID: 23718653DOI: 10.2460/ajvr.74.6.854Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research examined the effectiveness of innovative tools in extracting and concentrating bone marrow along with the separation of mesenchymal stem cells (MSC) in horses. The use of a unique gravitational marrow separator and a multidirectional needle were assessed in their capacity to collect and enhance quality of bone marrow. Furthermore, the research investigated the outcome of repeated bone marrow collections conducted a month apart.
Research Procedures and Methodology
- The study was conducted on eight healthy adult horses.
- Bone marrow collection was carried out twice, with a gap of one month in between, from the bodies of the animals using either a standard needle or a unique prototype multidirectional needle.
- The bone marrow obtained was concentrated using a gravitational marrow separator.
- Several aspects of the concentrated bone marrow were checked, including its counts of White Blood Cells (WBC) and platelets, automatic and cytomorphologic cell differential counts, and specifically MSCs, and cell viability.
Research Findings
- The bone marrow samples concentrated using the gravitational separator contained MSC, WBC, and platelet counts that were 5 to 19 times higher compared to unprocessed bone marrow samples.
- Utilization of the multidirectional needle enhanced the probability of deriving MSC-rich bone marrow.
- Repeating the bone marrow collection process after a month resulted in higher MSC numbers although there was a marginal decrease in cell viability post processing.
Conclusions and Clinical Applications
- The gravitational bone marrow separator and multidirectional needle proved effective in bone marrow extraction and enhancing quality of the concentrated bone marrow.
- Even higher or comparable amounts of bone marrow-derived MSCs were obtained when the aspiration operation was repeated a month later from the same harvesting sites.
- The gravity-based marrow separator and the multidirectional aspiration needle can provide a platform for a one-step, point-of-care, lab-independent method for acquiring concentrated bone marrow. This mixture comprises MSCs derived from bone marrow and growth factors from platelets and plasma.
Cite This Article
APA
Ishihara A, Helbig HJ, Sanchez-Hodge RB, Wellman ML, Landrigan MD, Bertone AL.
(2013).
Performance of a gravitational marrow separator, multidirectional bone marrow aspiration needle, and repeated bone marrow collections on the production of concentrated bone marrow and separation of mesenchymal stem cells in horses.
Am J Vet Res, 74(6), 854-863.
https://doi.org/10.2460/ajvr.74.6.854 Publication
Researcher Affiliations
- Comparative Orthopedic Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
MeSH Terms
- Animals
- Biopsy, Fine-Needle / instrumentation
- Biopsy, Fine-Needle / veterinary
- Bone Marrow Cells / cytology
- Bone Marrow Cells / physiology
- Cell Culture Techniques
- Cell Separation / instrumentation
- Cell Separation / methods
- Cell Separation / veterinary
- Female
- Horses
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / physiology
Citations
This article has been cited 4 times.- Liu Y, Zuo G, Meng X, Gao X, Zhang L, Tang P. Adrenomedullin inhibits osteoclast differentiation through the suppression of receptor activator of nuclear factor-κB ligand-induced nuclear factor-κB activation in glucocorticoid-induced osteoporosis. Exp Ther Med 2017 Nov;14(5):4009-4016.
- Bertone AL, Reisbig NA, Kilborne AH, Kaido M, Salmanzadeh N, Lovasz R, Sizemore JL, Scheuermann L, Kopp RJ, Zekas LJ, Brokken MT. Equine Dental Pulp Connective Tissue Particles Reduced Lameness in Horses in a Controlled Clinical Trial. Front Vet Sci 2017;4:31.
- Peters AE, Watts AE. Biopsy Needle Advancement during Bone Marrow Aspiration Increases Mesenchymal Stem Cell Concentration. Front Vet Sci 2016;3:23.
- Santiago-Torres JE, Lovasz R, Bertone AL. Fetal vs adult mesenchymal stem cells achieve greater gene expression, but less osteoinduction. World J Stem Cells 2015 Jan 26;7(1):223-34.
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