Neumann Katja, Dehne Tilo, Endres Michaela, Erggelet Christoph, Kaps Christian, Ringe Jochen, Sittinger Michael
TransTissue Technologies GmbH, Tucholskystrasse 2, 10117 Berlin, Germany.
J Orthop Res. 2008 Nov;26(11):1449-56. doi: 10.1002/jor.20635.
Microfracture is frequently used to repair articular cartilage defects and allows mesenchymal progenitors to migrate from subchondral bone into the defect and form cartilaginous repair tissue. The aim of our study was to analyze the cell surface antigen pattern and the differentiation capacity of cells derived from human subchondral bone. Human progenitor cells were derived from subchondral cortico-spongious bone and grown in the presence of human serum. Stem cell-related cell surface antigens were analyzed by flowcytometry. Cortico-spongious progenitor (CSP) cells showed presence of CD73, CD90, CD105, and STRO-1. Multilineage differentiation potential of CSP cells was documented by histological staining and by gene expression analysis of osteogenic, adipogenic, and chondrogenic marker genes. CSP cells formed a mineralized matrix as demonstrated by von Kossa staining and showed induction of osteocalcin, independent of osteogenic stimulation. During adipogenic differentiation, the adipogenic marker genes fatty acid binding protein 4 and peroxisome proliferative activated receptor gamma were induced. Immunohistochemical staining of cartilage-specific type II collagen and induction of the chondrocytic marker genes cartilage oligomeric matrix protein, aggrecan, and types II and IX collagen confirmed TGF beta 3-mediated chondrogenic lineage development. CSP cells from subchondral bone, as known from microfracture, are multipotent stem cell-like mesenchymal progenitors with a high chondrogenic differentiation potential.
微骨折术常用于修复关节软骨缺损,可使间充质祖细胞从软骨下骨迁移至缺损处并形成软骨修复组织。本研究旨在分析源自人软骨下骨的细胞的细胞表面抗原模式及分化能力。人祖细胞源自软骨下皮质海绵骨,并在人血清存在的条件下培养。通过流式细胞术分析干细胞相关的细胞表面抗原。皮质海绵祖细胞(CSP细胞)表达CD73、CD90、CD105和STRO-1。通过组织学染色以及对成骨、成脂和成软骨标记基因的基因表达分析,证明了CSP细胞具有多向分化潜能。如通过冯·科萨染色所示,CSP细胞形成了矿化基质,并显示出骨钙素的诱导表达,且不依赖于成骨刺激。在成脂分化过程中,诱导了成脂标记基因脂肪酸结合蛋白4和过氧化物酶体增殖物激活受体γ的表达。软骨特异性II型胶原蛋白的免疫组织化学染色以及软骨细胞标记基因软骨寡聚基质蛋白、聚集蛋白聚糖以及II型和IX型胶原蛋白的诱导表达,证实了转化生长因子β3介导的软骨细胞系发育。如微骨折术中所知,源自软骨下骨的CSP细胞是具有高软骨分化潜能的多能干细胞样间充质祖细胞。