Bagge Jasmin, MacLeod James N, Berg Lise C
Department of Veterinary Clinical Sciences, University of Copenhagen, Copenhagen, Denmark.
Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, KY, United States.
Front Vet Sci. 2020 Dec 10;7:602403. doi: 10.3389/fvets.2020.602403. eCollection 2020.
Bone marrow (BM)- and adipose tissue (AT)-derived mesenchymal stem cells (MSCs) are used increasingly for autologous cell therapy in equine practice to treat musculoskeletal and other injuries. Current recommendations often call for 10-100 million MSCs per treatment, necessitating the expansion of primary cells in culture prior to therapeutic use. Of concern, human and rodent studies have shown a decline of both MSC recovery from sampled tissue and proliferative capacity with increasing donor age. This may be problematic for applications of autologous cell-based therapies in the important equine demographic of older patients. To investigate the effect of donor age on the cellular proliferation of equine BM- and AT-MSCs. study. BM- and AT-MSCs and dermal fibroblasts (biological control) were harvested from horses in five different age groups ( = 4, = 60); newborn (0 days), yearling (15-17 months), adult (5-8 years), middle-aged (12-18 years), and geriatric (≥22 years). Proliferation of the cells was tested using an EdU incorporation assay and steady state mRNA levels measured for targeted proliferation, aging, and senescence biomarkers. The cellular proliferation of equine BM- and AT-MSCs declined significantly in the geriatric cohort relative to the younger age groups. Proliferation levels in the two MSC types were equally affected by donor age. Analysis of steady state mRNA levels showed an up-regulation in tumor suppressors, apoptotic genes, and multiple growth factors in MSCs from old horses, and a down-regulation of some pro-cycling genes with a few differences between cell types. Potential age-dependent differences in cell function parameters relevant to cell-therapy application were not investigated. The cellular proliferation of equine BM- and AT-MSCs declined at advanced donor ages. High levels of proliferation were observed in both MSC types from horses in the age groups below 18 years of age.
骨髓(BM)和脂肪组织(AT)来源的间充质干细胞(MSCs)在马医学实践中越来越多地用于自体细胞治疗,以治疗肌肉骨骼损伤和其他损伤。目前的建议通常要求每次治疗使用1亿至1亿个间充质干细胞,因此在治疗使用前需要在培养中扩增原代细胞。令人担忧的是,人类和啮齿动物研究表明,随着供体年龄的增加,从采样组织中回收的间充质干细胞和增殖能力都会下降。这对于在老年马匹这一重要群体中应用自体细胞疗法可能是个问题。为了研究供体年龄对马骨髓和脂肪组织间充质干细胞细胞增殖的影响。从五个不同年龄组(n = 4,n = 60)的马中采集骨髓和脂肪组织间充质干细胞以及真皮成纤维细胞(生物学对照);新生马(0天)、一岁马(15 - 17个月)、成年马(5 - 8岁)、中年马(12 - 18岁)和老年马(≥22岁)。使用EdU掺入试验检测细胞增殖,并测量靶向增殖、衰老和衰老生物标志物的稳态mRNA水平。与年轻年龄组相比,老年队列中马骨髓和脂肪组织间充质干细胞的细胞增殖显著下降。两种间充质干细胞类型的增殖水平受供体年龄的影响相同。稳态mRNA水平分析显示,老年马的间充质干细胞中肿瘤抑制因子、凋亡基因和多种生长因子上调,一些促细胞周期基因下调,细胞类型之间存在一些差异。未研究与细胞治疗应用相关的细胞功能参数中潜在的年龄依赖性差异。马骨髓和脂肪组织间充质干细胞的细胞增殖在供体年龄较大时下降。在18岁以下年龄组的马的两种间充质干细胞类型中均观察到高水平的增殖。