Peripheral mononuclear blood cell apheresis in a preclinical ovine model

Abstract

BackgroundRecent 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.ResultsSubcutaneous 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 × 106/ml to 16.1 ± 5.0 × 106/ml. Following apheresis, the WBC numbers in the apheretic product increased to 38.5 ± 27.6 × 106/ml, comprised of a significant increase in neutrophils and PBMC (from 5.25 ± 1.8 × 106/ml following G-CSF stimulation to 27.5 5 ± 27.6 × 106/ml). There was a mean of 2.1% CD34 + ve cells and 95.5% CD45 + ve cells in the apheretic product.ConclusionsThis 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.

Evaluation of the Safety and Feasibility of Apheresis in Dogs: For Application in Metastatic Cancer Research

“…At the connection between the CVC and the apheresis machine, which is the blood removal route and the extracorporeal circulation circuit, the formation of a thrombus along the circuit was observed. Blood coagulation and clots during apheresis have been observed in sheep [39] and dogs [24]. This major concern has been resolved by rigorous sterilization and continuous infusion of ACD-A solution.…”

Using apheresis‐derived cells to augment microdrilling in the treatment of chondral defects in an ovine model

“…Apheresis was performed under general anesthesia as described by Lydon et al 22 Catheters were placed into the jugular vein for blood collection and the cephalic vein for blood return. Apheresis was performed using a Cobe Spectra (Terumo‐BCT) with a disposable double‐needle closed system apheresis tubing kit.…”

<p>Role of Stem-Cell Transplantation in Leukemia Treatment</p>

“…A soft, gelatinous tissue, bone marrow is used as the source of peripheral HSCs. 69 Researchers have argued that both bone marrow and peripheral blood are major sources of SCs. SCASCs generated from bone marrow are known as bone-marrow SCs, 37 having clinical significance in restoring damaged cardiac tissue through gene therapy.…”

“… 74 PBSCs from bone marrow are a valuable source in restoring hematologic disorders. 69 The potential effect of PBSCs depends on hematopoietic development and enhancement of immunologic profiles, and hence they are a valuable source of HSCs to treat hematologic disorders. Peripheral blood CD34 + cells have hematopoietic potential for SCT.…”