Lang Hayley M, Schnabel Lauren V, Cassano Jennifer M, Fortier Lisa A
Department of Clinical Sciences, Cornell University, Ithaca, New York.
Vet Surg. 2017 Jul;46(5):731-737. doi: 10.1111/vsu.12639. Epub 2017 Mar 22.
Mesenchymal stem cells (MSCs) are frequently delivered via needle injection for treatment of musculoskeletal injuries. The purpose of this study was to evaluate the effect of needle diameter on the viability of MSCs.
Equine bone marrow-derived MSCs from 5 horses were suspended in PBS, and held at room temperature for 7 hours to mimic shipping conditions. Two replicate samples for each needle size (20, 22, 23, or 25-gauge [ga]) were aspirated into a 3 mL syringe and re-injected into the holding vial 3 times, to reproduce the resuspension of cells prior to injection in clinical cases. Cells were stained with fluorescein diacetate and propidium iodide to measure viability. Flow cytometry (FC) was performed to compare cell debris and intact cells between groups.
MSC viability was higher when cells were passed through a 20-ga rather than a 25-ga needle. Cell suspensions passed through a 20-ga needle contained a larger percentage of intact cells, compared to 25-ga samples. The percentage of debris present in cell suspensions tended to increase with decreasing needle diameter. Neither horse nor passage had a significant effect on viability.
Cell damage is more likely when MSCs are passed through 25-ga rather than 20-ga needles.
Use of needles larger than 25-ga is recommended to maintain the viability of MSCs injected in horses.
间充质干细胞(MSCs)常通过针剂注射来治疗肌肉骨骼损伤。本研究的目的是评估针的直径对MSCs活力的影响。
从5匹马获取的马骨髓源MSCs悬浮于磷酸盐缓冲液(PBS)中,在室温下保存7小时以模拟运输条件。将每种针尺寸(20、22、23或25号[ga])的两个重复样本吸入3毫升注射器中,并重新注入保存瓶3次,以重现临床病例中注射前细胞的重悬过程。用二乙酸荧光素和碘化丙啶对细胞进行染色以测量活力。进行流式细胞术(FC)以比较各组之间的细胞碎片和完整细胞。
当细胞通过20号针而非25号针时,MSCs活力更高。与25号样本相比,通过20号针的细胞悬液中完整细胞的百分比更高。细胞悬液中存在的碎片百分比倾向于随着针直径的减小而增加。马和传代次数对活力均无显著影响。
当MSCs通过25号针而非20号针时,细胞更易受损。
建议使用大于25号的针以维持注射到马体内的MSCs的活力。