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Peripheral mononuclear blood cell apheresis in a preclinical ovine model.

Published version
Peer-reviewed

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Authors

Lydon, Helen 
Brooks, Roger 

Abstract

Background. Recent research has demonstrated that circulating peripheral blood mononuclear fractions (PBMC) containing haematopoietic stem (HSC)/progenitor cells have the potential to play a crucial role in regenerative medicine strategies. Work in our laboratory has shown that a peripheral blood mononuclear cell fraction (PBMC) enhances cartilage repair in an osteochondral defect model in sheep and has a significant effect on cells in the joint niche. In order to obtain PBMC rich blood containing HSCs for further studies, we have performed, for the first time, apheresis on adult sheep. Results. Subcutaneous granulocyte-colony stimulating factor (G-CSF) was used to mobilise white blood cells and continual flow apheresis was performed on 8 sheep under general anaesthetic. There were no observable side effects, although a marked tendency for blood clotting during the procedure was noted. The administration of G-CSF for 3 days increased the white blood cell (WBC) count in the peripheral blood from to 6.7 ± 2.1 x 106/ml to 16.1 ± 5.0 x 106/ml. Following apheresis, the WBC numbers in the apheretic product increased to 38.5 ± 27.6 x 106/ml, comprised of a significant increase in neutrophils and PBMC (from 5.25 ± 1.8 x 106/ml following G-CSF stimulation to 27.5 5 ± 27.6 x 106/ml). There was a mean of 2.1% CD34+ve cells and 95.5% CD45+ve cells in the apheretic product Conclusions. This study describes the administration of G-CSF and subsequent apheresis in adult sheep. The technique is safe when performed as described with no observable side effects. The technique permits collection of an increased WBC fraction containing neutrophils and PBMC in adult sheep. This apheretic product contains CD34+ve cells, representing an HSC/progenitor population for use in in vivo and in vitro experiments.

Description

Keywords

Leukocytes, Mononuclear, Animals, Sheep, Granulocyte Colony-Stimulating Factor, Leukocyte Count, Leukapheresis, Female

Journal Title

BMC veterinary research

Conference Name

Journal ISSN

1746-6148
1746-6148

Volume Title

14

Publisher

BioMed Central
Sponsorship
ARTHRITIS RESEARCH UK (21156)
MRC (MC_PC_12009)