How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources

Musculoskeletal injuries in horses are very frequent and lead to substantial financial losses. Bone marrow mononuclear cells (BMMNCs), the fraction of bone marrow containing mesenchymal stem cells, have been successfully employed to enhance healing of tendon, ligament and articular injuries in this...

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Autores principales:	Corrêa,F, Borlone,C, Wittwer,F, Bustamante,H, Müller,A, Ramírez,A, Menarim,B
Lenguaje:	English
Publicado:	Facultad de Ciencias Veterinarias, Universidad Austral de Chile 2014
Materias:	cell separation horse musculoskeletal regenerative medicine
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2014000300019
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spelling	oai:scielo:S0301-732X20140003000192015-01-14How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resourcesCorrêa,FBorlone,CWittwer,FBustamante,HMüller,ARamírez,AMenarim,B cell separation horse musculoskeletal regenerative medicine Musculoskeletal injuries in horses are very frequent and lead to substantial financial losses. Bone marrow mononuclear cells (BMMNCs), the fraction of bone marrow containing mesenchymal stem cells, have been successfully employed to enhance healing of tendon, ligament and articular injuries in this specie. Many descriptions of how to harvest equine BM and isolate BMMNCs have been described; however, reported protocols consider fully equipped and costly laboratory facilities. The aim of this manuscript is to demonstrate to veterinary practitioners that obtaining and effective isolating equine BMMNCs for clinical use is possible with basic laboratory equipment. BM samples of 10 horses were collected from the 5th sternebra into 60 mL syringes containing heparin and Dulbecco's Phosphate-Buffered Saline (DPBS) using equipment that can be found in most equine clinics. Further, samples were centrifuged by density gradient for BMMNCs isolation within 1h after collection. Concentration and cell viability was determined by both direct counting on a Neubauer® chamber and epifluorescence analysis. Harvesting and isolation was achieved for all horses with no restrictions. The final 1 mL concentrate considered total number of BMMNCs (mean ± SD) and percentage of cell viability amongst isolates were 190±54 x 10(6) and 98±1.2%, respectively. Contamination was not observed in obtained samples and horses treated with BMMNCs using this technique did not present any clinical complications. Aspiration and processing of equine sternal BM using the described technique is a low cost and reliable method to obtain BMMNCs for cell based therapies.info:eu-repo/semantics/openAccessFacultad de Ciencias Veterinarias, Universidad Austral de ChileArchivos de medicina veterinaria v.46 n.3 20142014-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2014000300019en10.4067/S0301-732X2014000300019
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cell separation horse musculoskeletal regenerative medicine
spellingShingle	cell separation horse musculoskeletal regenerative medicine Corrêa,F Borlone,C Wittwer,F Bustamante,H Müller,A Ramírez,A Menarim,B How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
description	Musculoskeletal injuries in horses are very frequent and lead to substantial financial losses. Bone marrow mononuclear cells (BMMNCs), the fraction of bone marrow containing mesenchymal stem cells, have been successfully employed to enhance healing of tendon, ligament and articular injuries in this specie. Many descriptions of how to harvest equine BM and isolate BMMNCs have been described; however, reported protocols consider fully equipped and costly laboratory facilities. The aim of this manuscript is to demonstrate to veterinary practitioners that obtaining and effective isolating equine BMMNCs for clinical use is possible with basic laboratory equipment. BM samples of 10 horses were collected from the 5th sternebra into 60 mL syringes containing heparin and Dulbecco's Phosphate-Buffered Saline (DPBS) using equipment that can be found in most equine clinics. Further, samples were centrifuged by density gradient for BMMNCs isolation within 1h after collection. Concentration and cell viability was determined by both direct counting on a Neubauer® chamber and epifluorescence analysis. Harvesting and isolation was achieved for all horses with no restrictions. The final 1 mL concentrate considered total number of BMMNCs (mean ± SD) and percentage of cell viability amongst isolates were 190±54 x 10(6) and 98±1.2%, respectively. Contamination was not observed in obtained samples and horses treated with BMMNCs using this technique did not present any clinical complications. Aspiration and processing of equine sternal BM using the described technique is a low cost and reliable method to obtain BMMNCs for cell based therapies.
author	Corrêa,F Borlone,C Wittwer,F Bustamante,H Müller,A Ramírez,A Menarim,B
author_facet	Corrêa,F Borlone,C Wittwer,F Bustamante,H Müller,A Ramírez,A Menarim,B
author_sort	Corrêa,F
title	How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
title_short	How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
title_full	How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
title_fullStr	How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
title_full_unstemmed	How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
title_sort	how to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources
publisher	Facultad de Ciencias Veterinarias, Universidad Austral de Chile
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0301-732X2014000300019
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How to obtain and isolate equine sternal bone marrow mononuclear cells with limited resources

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