Characterization of canine mesenchymal stromal cell lines (MSC)

The bone marrow microenvironment is a complex mixture of cells that function in concert to regulate hematopoiesis. Cellular components include fixed nonhematopoietic stromal elements (MSC) as well as monocytes and resident macrophages, which are derived from the hematopoietic stem cells. Clinical studies have shown healing effects, direct or indirect, from the injection of MSC int Keywords: Cell type comparison Canine bone marrow stromal cells were immortalized by transduction with a replication-defective recombinant retrovirus containing the human papilloma virus E6/E7 genes (pLXSN-16 E6E7), as described for human stromal cell lines in Roecklein and Torok-Storb (Blood 85:997-1005, 1995). Transduced clones were selected with 50 ug/ml G418 and resistant clones were isolated, grown to confluency and characterized for morphology, cytokine production and cell surface molecule expression. These cells are intended for clinical research in the canine transplantation model, as a complement for existing human stromal cell lines, for mitigation of graft-versus-host disease and radiation effects.

骨髓微环境是一类由多种细胞共同构成的复杂微环境，各类细胞协同发挥功能以调控造血过程。该微环境的细胞组分包括固定的非造血基质成分（间充质干细胞，Mesenchymal Stem Cell, MSC）、单核细胞以及源自造血干细胞的驻留巨噬细胞。临床研究表明，间充质干细胞注射可产生直接或间接的修复效应。
关键词：细胞类型比较
本研究通过携带人乳头瘤病毒E6/E7基因的复制缺陷型重组逆转录病毒（pLXSN-16 E6E7）对犬骨髓基质细胞进行转导以实现永生化，具体方法参照Roecklein与Torok-Storb针对人源基质细胞系的报道（Blood, 1995, 85:997-1005）。转导后的克隆通过50 μg/ml G418进行筛选，分离得到抗性克隆；待抗性克隆生长至汇合状态后，分别对其形态学特征、细胞因子分泌能力以及细胞表面分子表达情况进行鉴定。本永生化细胞系旨在用于犬移植模型的临床研究，作为现有人源基质细胞系的补充工具，用于缓解移植物抗宿主病以及辐射相关损伤效应。