Cryopreserved CD90+ cells obtained from mobilized peripheral blood in sheep: a new source of mesenchymal stem cells for preclinical applications

Abstract

Mobilized peripheral blood (MPB) bone marrow cells possess the potential to differentiate into a variety of mesenchymal tissue types and offer a source of easy access for obtaining stem cells for the development of experimental models with applications in tissue engineering. In the present work, we aimed to isolate by magnetic activated cell sorting CD90+ cells from MPB by means of the administration of Granulocyte-Colony Stimulating Factor and to evaluate cell proliferation capacity, after thawing of the in vitro culture of this population of mesenchymal stem cells (MSCs) in sheep. We obtained a median of 8.2 ± 0.6 million of CD90+ cells from the 20-mL MPB sample. After thawing, at day 15 under in vitro culture, the mean CD90+ cells determined by flow cytometry was 92.92 ± 1.29 % and cell duplication time determined by crystal violet staining was 47.59 h. This study describes for the first time the isolation, characterization, and post-in vitro culture thawing of CD90+ MSCs from mobilized peripheral blood in sheep. This population can be considered as a source of MSCs for experimental models in tissue engineering research.

Ovine Mesenchymal Stromal Cells: Morphologic, Phenotypic and Functional Characterization for Osteochondral Tissue Engineering

“…Previous studies have reported the isolation of oMSCs from bone marrow [ 5 , 23 – 25 ], adipose tissue [ 34 – 36 ], umbilical cord blood [ 36 , 37 ], peripheral blood [ 21 , 38 ], amniotic fluid [ 39 , 40 ], dermis [ 41 ] and periodontal ligament [ 42 ]. Despite the increasing use of sheep as a large animal model for tissue engineering, because of their similarities to humans in size, joint architecture and healing mechanisms [ 25 ], ovine MSCs have been poorly characterized compared to human MSCs [ 24 ].…”

Monosodium urate crystals induce oxidative stress in human synoviocytes

“…To evaluate surface markers associated with fibroblasts and macrophage, a flow cytometry (FC) assay was performed according to Landa-Solís C et al [ 16 ] Cells were marked with monoclonal antibodies PE-conjugated CD166 and PE-conjugated CD14 from BD PharMigenTM (San Diego, CA, USA). Data were collected through a BD FACSCalibur flow cytometer and analyzed with CellQuestTM PRO software (Becton-Dickinson).…”

Effects of tacrolimus on morphology, proliferation and differentiation of mesenchymal stem cells derived from gingiva tissue

“…Mesenchymal stem cells (MSCs) are multipotent progenitor cells that have the capacity for self-renewal and differentiation into osteocytes, adipocytes and other cells ( 1 ). MSCs have been used in preclinical models for tissue engineering of bone, cartilage, muscle, marrow stroma, tendon, fat and other connective tissues ( 2 – 4 ). In addition, MSCs secrete a broad spectrum of bioactive macromolecules that are immunomodulatory and serve to structure regenerative microenvironments in fields of tissue injury ( 5 ).…”