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Evaluation of the Cell Viability of Human Whartons Jelly Stem Cells for Use in Cell Therapy

机译:沃顿商学院果冻干细胞在细胞治疗中的细胞活力评估

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摘要

Human umbilical cord Wharton's jelly stem cells (HWJSCs) are gaining attention as a possible clinical source of mesenchymal stem cells for cell therapy and tissue engineering due to their high accessibility, expansion potential, and plasticity. We employed a combination of highly sensitive techniques to determine the average cell viability levels and proliferation capabilities of 10 consecutive cell passages of cultured HWJSCs and then used RNA microarrays to identify genes associated with changes in cell viability levels. We found an initial decrease in cell viability from the first to the third cell passage followed by an increase until the sixth passage and a final decrease from the sixth to tenth cell passages. The highest cell viability levels corresponded to the fifth and sixth passages. The intracellular ionic contents of potassium, sodium, and chlorine suggest that the lower cell viability levels at passages 2, 3, and 8–10 may be associated with apoptotic cell death. In fact, gene expression analysis revealed that the average cell viability was significantly associated with genes with a function in apoptotic cell death, especially pro-apoptotic FASTKD2, BNIP3L genes and anti-apoptotic TNFAIP8 and BCL2L2 genes. This correlation with both pro-apoptotic and anti-apoptotic genes suggests that there may be a complex live-death equilibrium in cultured HWJSCs kept in culture for multiple cell passages. In this study, the highest cell viability levels corresponded to the fifth and sixth HWJSC passages, suggesting that these passages should be preferentially employed in cell therapy or tissue engineering protocols using this cell type.
机译:人脐带沃顿氏胶冻干细胞(HWJSCs)由于其高可及性,可扩展性和可塑性而成为可能用于细胞治疗和组织工程的间充质干细胞的临床来源。我们采用了高灵敏度技术的组合来确定培养的HWJSC的10个连续细胞传代的平均细胞活力水平和增殖能力,然后使用RNA微阵列来鉴定与细胞活力水平变化相关的基因。我们发现细胞活力从第一个到第三次传代开始下降,然后一直上升到第六个传代,最后从第六到第十次传代下降。最高细胞活力水平对应于第五和第六代。钾,钠和氯的细胞内离子含量表明,第2、3和8-10代较低的细胞活力水平可能与凋亡细胞死亡有关。实际上,基因表达分析表明,平均细胞活力与具有凋亡细胞死亡功能的基因显着相关,尤其是促凋亡的FASTKD2,BNIP3L基因以及抗凋亡的TNFAIP8和BCL2L2基因。与促凋亡和抗凋亡基因的这种相关性表明,在培养物中多次培养的HWJSC中可能存在复杂的活死平衡。在这项研究中,最高的细胞活力水平对应于第五和第六次HWJSC传代,表明这些传代应优先用于使用这种细胞类型的细胞疗法或组织工程方案中。

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