Pattappa Girish, Reischl Franziska, Jahns Judith, Schewior Ruth, Lang Siegmund, Zellner Johannes, Johnstone Brian, Docheva Denitsa, Angele Peter
Laboratory for Experimental Trauma Surgery, Department of Trauma Surgery, University Regensburg Medical Centre, Regensburg, Germany.
Sporthopaedicum Regensburg, Regensburg, Germany.
Front Bioeng Biotechnol. 2022 Jan 28;9:789621. doi: 10.3389/fbioe.2021.789621. eCollection 2021.
The meniscus is composed of an avascular inner region and vascular outer region. The vascular region has been shown to contain a progenitor population with multilineage differentiation capacity. Strategies facilitating the isolation and propagation of these progenitors can be used to develop cell-based meniscal therapies. Differential adhesion to fibronectin has been used to isolate progenitor populations from cartilage, while low oxygen or physioxia (2% oxygen) enhances the meniscal phenotype. This study aimed to isolate progenitor populations from the avascular and vascular meniscus using differential fibronectin adherence and examine their clonogenicity and differentiation potential under hyperoxia (20% oxygen) and physioxia (2% oxygen). Human vascular and avascular meniscus cells were seeded onto fibronectin-coated dishes for a short period and monitored for colony formation under either hyperoxia or physioxia. Non-fibronectin adherent meniscus cells were also expanded under both oxygen tension. Individual fibronectin adherent colonies were isolated and further expanded, until approximately ten population doublings (passage 3), whereby they underwent chondrogenic, osteogenic, and adipogenic differentiation. Physioxia enhances clonogenicity of vascular and avascular meniscus cells on plastic or fibronectin-coated plates. Combined differential fibronectin adhesion and physioxia isolated a progenitor population from both meniscus regions with trilineage differentiation potential compared to equivalent hyperoxia progenitors. Physioxia isolated progenitors had a significantly enhanced meniscus matrix content without the presence of collagen X. These results demonstrate that combined physioxia and fibronectin adherence can isolate and propagate a meniscus progenitor population that can potentially be used to treat meniscal tears or defects.
半月板由无血管的内部区域和血管丰富的外部区域组成。已证明血管丰富区域含有具有多向分化能力的祖细胞群。促进这些祖细胞分离和增殖的策略可用于开发基于细胞的半月板治疗方法。利用对纤连蛋白的差异黏附从软骨中分离祖细胞群,而低氧或生理性低氧(2%氧气)可增强半月板表型。本研究旨在利用纤连蛋白差异黏附从无血管和血管丰富的半月板中分离祖细胞群,并检测它们在高氧(20%氧气)和生理性低氧(2%氧气)条件下的克隆形成能力和分化潜能。将人血管丰富和无血管的半月板细胞短期接种到纤连蛋白包被的培养皿上,并在高氧或生理性低氧条件下监测集落形成。非纤连蛋白黏附的半月板细胞也在两种氧张力条件下进行扩增。分离出单个纤连蛋白黏附的集落并进一步扩增,直至大约十个群体倍增(传代3次),然后进行软骨形成、成骨和成脂分化。生理性低氧增强了血管丰富和无血管半月板细胞在塑料或纤连蛋白包被平板上的克隆形成能力。与同等高氧祖细胞相比,纤连蛋白差异黏附与生理性低氧相结合从两个半月板区域分离出具有三系分化潜能的祖细胞群。生理性低氧分离出的祖细胞在不存在X型胶原的情况下半月板基质含量显著增加。这些结果表明,生理性低氧与纤连蛋白黏附相结合能够分离和扩增半月板祖细胞群,该细胞群有可能用于治疗半月板撕裂或缺损。