Kerlan Jobe Institute, Cedars-Sinai Medical Center, Los Angeles, California, U.S.A.
Orthopedic Stem Cell Research Laboratory, Cedars-Sinai Medical Center, Los Angeles, California, U.S.A.; Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, California, U.S.A.
Arthroscopy. 2021 Jan;37(1):252-265. doi: 10.1016/j.arthro.2020.09.021. Epub 2020 Sep 23.
To identify, characterize, and compare the resident progenitor cell populations within the red-red, red-white, and white-white (WW) zones of freshly harvested human cadaver menisci and to characterize the vascularity of human menisci using immunofluorescence and 3-dimensional (3D) imaging.
Fresh adult human menisci were harvested from healthy donors. Menisci were enzymatically digested, mononuclear cells isolated, and characterized using flow cytometry with antibodies against mesenchymal stem cell surface markers (CD105, CD90, CD44, and CD29). Cells were expanded in culture, characterized, and compared with bone marrow-derived mesenchymal stem cells. Trilineage differentiation potential of cultured cells was determined. Vasculature of menisci was mapped in 3D using a modified uDisco clearing and immunofluorescence against vascular markers CD31, lectin, and alpha smooth muscle actin.
There were no significant differences in the clonogenicity of isolated cells between the 3 zones. Flow cytometry showed presence of CD44CD105CD29CD90 cells in all 3 zones with high prevalence in the WW zone. Progenitors from all zones were found to be potent to differentiate to mesenchymal lineages. Larger vessels in the red-red zone of meniscus were observed spanning toward red-white, sprouting to smaller arterioles and venules. CD31 cells were identified in all zones using the 3D imaging and co-localization of additional markers of vasculature (lectin and alpha smooth muscle actin) was observed.
The presence of resident mesenchymal progenitors was evident in all 3 meniscal zones of healthy adult donors without injury. In addition, our results demonstrate the presence of vascularization in the WW zone.
The existence of progenitors and presence of microvasculature in the WW zone of the meniscus suggests the potential for repair and biologic augmentation strategies in that zone of the meniscus in young healthy adults. Further research is necessary to fully define the functionality of the meniscal blood supply and its implications for repair.
鉴定、描述并比较新鲜人尸体半月板的红白区和白白区(WW 区)内的常驻祖细胞群体,并通过免疫荧光和 3 维(3D)成像来描述人半月板的血管分布。
从健康供体中收获新鲜成人半月板。半月板经酶消化,分离单核细胞,并用针对间充质干细胞表面标志物(CD105、CD90、CD44 和 CD29)的抗体进行流式细胞术进行鉴定。细胞在培养中进行扩增、鉴定,并与骨髓间充质干细胞进行比较。检测培养细胞的三系分化潜能。使用改良 uDisco 透明化和针对血管标志物 CD31、凝集素和α平滑肌肌动蛋白的免疫荧光在 3D 中描绘半月板的血管分布。
3 个区域分离细胞的集落形成能力无显著差异。流式细胞术显示,所有 3 个区域均存在 CD44CD105CD29CD90 细胞,WW 区域的阳性率较高。所有区域的祖细胞均被证明能够向间充质谱系分化。半月板红-红区较大的血管向红白区延伸,向较小的动静脉分支延伸。使用 3D 成像观察到所有区域均存在 CD31 细胞,并且观察到血管的其他标志物(凝集素和α平滑肌肌动蛋白)的共定位。
在没有损伤的健康成年供体的所有 3 个半月板区域均存在常驻间充质祖细胞。此外,我们的结果表明 WW 区存在血管化。
半月板 WW 区祖细胞的存在和微脉管系统的存在提示在年轻健康成年人的半月板 WW 区有修复和生物增强策略的潜力。需要进一步的研究来充分定义半月板血液供应的功能及其对修复的影响。