The effect of delivery via narrow-bore needles on mesenchymal cells
The effect of delivery via narrow-bore needles on mesenchymal cells
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DOI：
10.2217/17460751.4.1.49
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发表时间：
2009-01-01
影响因子：
2.7
通讯作者：
Shakesheff, Kevin M.
中科院分区：
文献类型：
--
作者：
Agashi, Kapil;Chau, David Y. S.;Shakesheff, Kevin M.
目的：最近，有许多临床前和人体研究，研究细胞悬浮液直接注射到靶器官后的再生能力：成功的(临床)细胞治疗的基本参数。目前，在这些细胞通过窄孔针排出过程中和之后，对这些细胞存活的重要因素(即形态、活力和增殖率)的识别数据有限。材料与方法：分离、扩增小鼠骨髓间充质干细胞(MMSCs)，制成含HBSS或HBSS加抗氧化剂N-乙酰半胱氨酸的浓缩细胞悬液。然后，通过不同口径的针头，以不同的射出速度从10亩的L汉密尔顿注射器中喷射出这种悬浮液。然后检测细胞活性、铺展和贴壁、细胞凋亡和增殖能力。结果：在注射器内操作后，mMSCs的活性和细胞铺展能力下降，同时caspase-3蛋白的产生增加，这是细胞凋亡的早期调节事件。这些有害影响随着细胞在注射器腔内放置时间的增加而增加，在5亩L/分钟和1亩L/分钟的射出速度下，这些有害影响相似。然而，随着针孔直径的增加，观察到这些特性显著降低。相比之下，在室温(18-20摄氏度)下放置，但没有在注射器中操作的mMSCs，显示出比操作细胞明显更高的活性。然而，保持在4摄氏度的细胞显示出与操纵细胞相比存活率降低。当MMSC与已知的抗氧化剂N-乙酰半胱氨酸孵育时，caspase-3的产生或细胞铺展没有显著变化。结论：这项研究强调了潜在的参数，如最小化细胞在注射器中的时间段和使用更大口径的针头，这些参数涉及到维持细胞治疗应用所需的细胞悬浮液输送所需的高活细胞密度。
Aims: Recently, there have been numerous preclinical and human studies investigating the regenerative capacity of cell suspensions following their direct injection into a target organ: the fundamental parameters for successful (clinical) cell therapy. At present, limited data exist in the identification of factors important for the survival of these cells (i.e., morphology, viability and proliferation rates) during and following their ejection via narrow-bore needles. Materials & methods: Primary murine mesenchymal stem cells (mMSCs) were isolated, expanded and processed into a concentrated cell suspension consisting of either HBSS or HBSS supplemented with the antioxidant n-acetyl-cysteine. This suspension was then ejected from a 10 mu l Hamilton syringe, via a variety of bore-sized needles, at different ejection rates. Cell characteristics including viability, spreading and attachment, apoptosis and proliferative ability were then assessed. Results: Following manipulation within a syringe, a decrease in the viability and cell spreading of mMSCs and a concurrent increase in the production of the caspase-3 protein, an early regulatory event in apoptosis, occurs. These detrimental effects were found to be increased when the cells were left in the syringe chamber for increased periods of time, and were similar at 5 mu l/min and 1 mu l/min ejection rates. However, on increasing the needle bore diameter, a significant reduction in these characteristics was observed. By comparison, mMSCs that were left to stand at room temperature (18-20 degrees C), but were not manipulated within a syringe, showed a significantly greater viability compared with manipulated cells. However, cells kept at 4 degrees C demonstrated a decreased viability compared with manipulated cells. When the mMSC were incubated with n-acetyl-cysteine, a known antioxidant, no significant change in caspase-3 production or cell spreading was observed. Conclusions: This study highlights potential parameters, such as minimizing the time period the cells are within the syringe and the use of wider-bore needles, involved in maintaining the high viable cell density required for the delivery of cell suspensions for cell therapy applications.