Chen Xia, Thibeault Susan
Annu Int Conf IEEE Eng Med Biol Soc. 2013;2013:6228-31. doi: 10.1109/EMBC.2013.6610976.
For cells seeded in scaffolds, transplanted cell survival rate plays an important role for cell transplantation efficiency, and is essential for successful cell transplantation. Fibroblast viability in HyStem-C was examined by a double staining Live/Dead Viability/Cytotoxicity assay, and cell images were analyzed using MetaMorph software for calculating live cell percentage for fresh and cryopreserved cells at different incubation time points, delivery methods, differing DMSO and cell concentrations. The results of this research demonstrated that in HyStem-C, the viability of cryopreserved cells (85%) was significantly lower than fresh collected cells (96.7%). In addition, the physical force from a 27 gauge needle significantly decreased frozen cell survival rates to 83-85% compared to pipette delivered cells. Higher DMSO concentration (1.0%) and higher cell density (2 × 10(7) per milliliter) also significantly decreased cell survival to 73%. Cryopreserved cell viability in three dimensional scaffolding can be maintained over 80% with cell density of 1 × 10(7) per milliliter, total DMSO concentration of 0.5%, and passed through a 27-gauge needle. These results demonstrate the viability of cells seeded in hyaluronan hydrogel with commonly used storage and delivery methods can bring rather satisfactory cell transplantation efficiency.
对于接种在支架中的细胞，移植细胞存活率对细胞移植效率起着重要作用，并且是成功进行细胞移植所必需的。通过活/死细胞活力/细胞毒性双染法检测了HyStem-C中培养的成纤维细胞的活力，并使用MetaMorph软件分析细胞图像，以计算不同孵育时间点、递送方法、不同二甲基亚砜（DMSO）和细胞浓度下新鲜细胞和冻存细胞的活细胞百分比。本研究结果表明，在HyStem-C中，冻存细胞的活力（85%）显著低于新鲜收集的细胞（96.7%）。此外，与通过移液器递送的细胞相比，27号针头产生的物理力使冻存细胞存活率显著降低至83 - 85%。较高的DMSO浓度（1.0%）和较高的细胞密度（每毫升2×10⁷个）也使细胞存活率显著降低至73%。当细胞密度为每毫升1×10⁷个、DMSO总浓度为0.5%并通过27号针头时，三维支架中冻存细胞的活力可维持在80%以上。这些结果表明，采用常用的储存和递送方法接种在透明质酸水凝胶中的细胞活力能够带来相当令人满意的细胞移植效率。
