Department of Histology-Tissue Engineering Group, University of Granada, Granada, Spain.
Tissue Eng Part C Methods. 2012 Jun;18(6):408-19. doi: 10.1089/ten.TEC.2011.0508. Epub 2012 Jan 26.
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.
人脐带华通氏胶干细胞(HWJSCs)由于其高易得性、扩增潜力和可塑性,正作为间充质干细胞的一种潜在临床来源而受到关注,可用于细胞治疗和组织工程。我们采用了一系列高灵敏度技术来确定培养的 HWJSCs 连续 10 代的细胞平均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代到第三代培养时会先下降,然后在第六代培养时会增加,最后在第六到第十代培养时会再次下降。细胞活力最高的对应第五代和第六代培养。钾、钠和氯的细胞内离子含量表明,在第 2、3 代和第 8-10 代时较低的细胞活力可能与细胞凋亡死亡有关。事实上,基因表达分析显示,细胞平均活力与凋亡细胞死亡功能相关的基因显著相关,特别是促凋亡 FASTKD2、BNIP3L 基因和抗凋亡 TNFAIP8 和 BCL2L2 基因。这种与促凋亡和抗凋亡基因的相关性表明,在经过多次传代培养的培养 HWJSCs 中,可能存在一个复杂的生死平衡。在这项研究中,细胞活力最高的对应于第五代和第六代 HWJSC 培养,这表明在使用这种细胞类型的细胞治疗或组织工程方案中,应优先选用这几代。