Processing of equine bone marrow using the automated MarrowXpress System: RBC depletion, volume reduction, and mononuclear cell recovery

Abstract

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.

Equine bone marrow volume reduction, red blood cell depletion, and mononuclear cell recovery using the PrepaCyte‐CB processing system

“…This study shows that adequate reduction in RBC number and sample volume as well as enhanced recovery of MNC, presumably containing MSC, can be achieved by processing equine BM with the PrepaCyte‐CB system. In addition, MNC recovery using the PrepaCyte‐CB processing system was higher than that reported for equine BM using an automated system, with superior RBC and volume depletion (Table ). RBC and volume depletion in the current study were both less than, but comparable, to that described for human umbilical cord blood processed by the PrepaCyte‐CB system (Table ) .…”

“…In human studies, the PrepaCyte‐CB system provides fast and reliable blood separation of human cord blood, with superior recovery of nucleated cells and removal of RBC compared to another standard blood separation method using hydroxyethyl starch . It also requires less equipment and technical expertise than automated processing systems currently used in veterinary medicine …”

“…Protocols for isolation of equine BM MNCs are rarely reported in the literature and describe 2 main standard methods of cell separation . The first method uses density gradient solutions such as Ficoll‐Paque Plus (GE Healthcare Life Sciences, Pittsburgh, PA, USA) to separate BM‐MNC fraction from other blood cells; however, volume reduction is not achieved .…”

“…The first method uses density gradient solutions such as Ficoll‐Paque Plus (GE Healthcare Life Sciences, Pittsburgh, PA, USA) to separate BM‐MNC fraction from other blood cells; however, volume reduction is not achieved . The second method utilizes an automated system such as the MarrowXpress System (CESCA Therapeutics, Rancho Cordova, CA, USA) to separate MNCs while depleting RBC number and reducing sample volume …”

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

“…Both have demonstrated to enhance the repair of soft tissue injuries in horses (Smith et al 2003, Oliveira 2008, Crovace et al 2010, nonetheless, all the described protocols require access to fully equipped and costly laboratory facilities. Besides, obtaining a pure fraction of cultivated MSCs also demands longer time (2-3 weeks) (Fortier andSmith 2008, Kol et al 2013) and higher costs related to cell culture (Owens et al 2011). This study aims to demonstrate to equine and other veterinary practitioners that obtaining and isolating BMMNCs from horses is a simple procedure and can be performed with basic laboratory equipment.…”