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response in vivo and, therefore, improved tendon tissue healing. Therefore, the objectives of this study were to 1) determine the MSC cell surface expression of MHC class I and transcriptome-wide gene expression changes following IL-1β + TGF-β2 dual

Open access
in Journal of the American Veterinary Medical Association

Abstract

OBJECTIVE To assess efficiency of gravity filtration to enhance recovery of equine bone marrow elements including stem and progenitor cells.

ANIMALS 12 healthy adult horses.

PROCEDURES Bone marrow aspirates were collected from the fifth sternebral body and filtered by gravitational flow to obtain bone marrow elements. Raw and harvested bone marrow and marrow effluent were evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, mesenchymal stem cell CFUs, cell viability, and differentiation capacity. Isolated cells were analyzed for CD90 and major histocompatibility complex (MHC) class I and II antigens.

RESULTS Mean cell viability of harvested bone marrow was 95.9%. Total WBCs and platelets were efficiently captured on the filter (> 95%), and mean recovery in harvested bone marrow was 30%. Cytologic cell differential counts indicated that the percentage of neutrophils was significantly less and the progenitor cell population was significantly higher and concentrated 1.56-fold in harvested bone marrow, compared with results for raw bone marrow. Flow cytometry and cell culture were used to characterize harvested bone marrow cells as positive for expression of CD90 and negative for MHCI and MHCII, which indicated stem cells with a multipotent phenotype that differentiated into chondrocytes, osteocytes, adipocytes, and tenocytes.

CONCLUSIONS AND CLINICAL RELEVANCE Gravitational filtration of bone marrow efficiently yielded platelets and cells and produced a progenitor-enriched, leukocyte-reduced product, compared with raw bone marrow.

Full access
in American Journal of Veterinary Research

assigned medium was replaced 48 hours after initiation of incubation to adequately maintain the MSCs that were collected after 96 hours of incubation. Expression of MHC class I and MHC class II was analyzed by flow cytometry as described. 13 For the qRT

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in American Journal of Veterinary Research

selected for qRT-PCR assay on the basis of the 5 genes representing the most significant expression changes as determined with the microarray. These genes included those for IgC, MHC class II HLA-DRB1, CCL5, MHC class I DLA-64, and MHC class I DLA-88. Gene

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in American Journal of Veterinary Research

has 2 major functions. First, TGF-β inhibits interferon γ–induced MHC gene expression. Subsequently, downregulation of MHC class I allows tumor cells to escape recognition and destruction by cytotoxic T lymphocytes and natural killer cells. Moreover

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in Journal of the American Veterinary Medical Association

polymorphic microsatellite marker within the canine MHC class I region . Mamm Genome 1995 ; 6 : 684 – 685 . 7 Mellersh CS Langston AA Acland GM , et al . A linkage map of the canine genome . Genomics

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in Journal of the American Veterinary Medical Association

that recognize MHC class I molecules. 3,4 Those T helper cells have an important role in the adaptive immune system by orchestrating a cascade of reactions resulting in secretion of proinflammatory cytokines and cytotoxic molecules and the expression

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in American Journal of Veterinary Research

an important marker for macrophage identification. 17 Moreover, activated macrophages also express the MHC, which is an important immune molecule that has a role in presenting endogenous and exogenous antigens. 26 The MHC class I and II molecules

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in American Journal of Veterinary Research

A , et al. A cis-acting regulatory mutation causes premature hair graying and susceptibility to melanoma in the horse . Nat Genet 2008 ; 40 : 1004 – 1009 . 10.1038/ng.185 12. Seliger B . Molecular mechanisms of MHC class I abnormalities and

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in American Journal of Veterinary Research

) lymphoid markers and myeloid markers (CD14, clone 105; CD172a [clone DH59B]) 1–4 but did express MHC class I (clone cz3.1) and MHC class II (clone cz11) markers. 2,3 Immunocytochemical staining of peripheral blood and bone marrow aspirate smears revealed

Full access
in Journal of the American Veterinary Medical Association