Ganguly Payal, El-Jawhari Jehan J, Burska Agata N, Ponchel Frederique, Giannoudis Peter V, Jones Elena A
Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, UK.
Department of Clinical Pathology, Faculty of Medicine, Mansoura University, Egypt.
Stem Cells Int. 2019 Aug 1;2019:5197983. doi: 10.1155/2019/5197983. eCollection 2019.
Uncultured mesenchymal stromal cells (MSCs) are increasingly used in therapies; however, the effects of donor age on their biological characteristics and gene expression remain unclear. The aim of this study was to investigate age-related changes in bone marrow (BM) MSCs following minimal or no culture manipulation. Iliac crest BM was aspirated from 67 healthy donors (19-89 years old) and directly used for the colony-forming unit-fibroblast (CFU-F) assay or CD45CD271 cell enumeration. The colonies were analysed for colony area and integrated density (ID) when grown in standard MSC media or media supplemented with human serum from young (YS) or old (OS) donors. There was a notable age-related decline in the number of MSCs per millilitre of BM aspirate revealed by the CFU-F assay ( = -0.527, < 0.0001) or flow cytometry ( = -0.307, = 0.0116). Compared to young donors (19-40 years old), colony IDs were significantly lower in older donors (61-89 years old), particularly for smaller-sized colonies (42% lower, < 0.01). When cultured in media supplemented with OS, young and old donor MSCs formed colonies with lower IDs, by 21%, < 0.0001, and 27%, < 0.05, respectively, indicating the formation of smaller sparser colonies. No significant differences in the expression of selected adipogenic, osteogenic, stromal, and bone remodelling genes as well as CD295, CD146, CD106, and connexin 43 surface molecules were found in sorted CD45CD271 MSCs from young and old donors ( = 8 donors each). Altogether, these results show similar trends for age-related decline in BM MSC numbers measured by the CFU-F assay and flow cytometry and reveal age-related effects of human serum on MSC colony formation. No significant differences in selected gene expression in uncultured CD45CD271 MSCs suggest that old donor MSCs may not be inferior in regard to their multipotential functions. Due to large donor-to-donor variation in all donor groups, our data indicate that an individual's chronological age is not a reliable predictor of their MSC number or potency.
未培养的间充质基质细胞（MSCs）越来越多地用于治疗；然而，供体年龄对其生物学特性和基因表达的影响仍不清楚。本研究的目的是调查在极少或无培养操作的情况下，骨髓（BM）间充质干细胞随年龄的变化。从67名健康供体（19 - 89岁）中抽取髂嵴骨髓，直接用于集落形成单位-成纤维细胞（CFU-F）测定或CD45⁻CD271⁺细胞计数。当在标准间充质干细胞培养基或补充了年轻（YS）或年老（OS）供体人血清的培养基中生长时，分析集落的面积和积分密度（ID）。CFU-F测定（r = -0.527，P < 0.0001）或流式细胞术（r = -0.307，P = 0.0116）显示，每毫升骨髓抽吸物中间充质干细胞的数量随年龄有显著下降。与年轻供体（19 - 40岁）相比，年老供体（61 - 89岁）的集落ID显著更低，尤其是对于较小尺寸的集落（低42%，P < 0.01）。当在补充了OS的培养基中培养时，年轻和年老供体的间充质干细胞形成的集落ID分别降低21%（P < 0.0001）和27%（P < 0.05），表明形成了更小、更稀疏的集落。在来自年轻和年老供体的分选CD45⁻CD271⁺间充质干细胞中（每组n = 8名供体），未发现所选的脂肪生成、成骨、基质和骨重塑基因以及CD295、CD146、CD106和连接蛋白43表面分子的表达有显著差异。总之，这些结果表明，CFU-F测定和流式细胞术测量的骨髓间充质干细胞数量随年龄下降呈现相似趋势，并揭示了人血清对间充质干细胞集落形成的年龄相关影响。未培养的CD45⁻CD271⁺间充质干细胞所选基因表达无显著差异，表明年老供体的间充质干细胞在多能功能方面可能并不逊色。由于所有供体组中供体间差异较大，我们的数据表明，个体的实际年龄并非其间充质干细胞数量或潜能的可靠预测指标。
