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Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes

机译:脐部单核细胞和成纤维细胞相互作用下调细胞周期阴性对照基因的表达

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摘要

The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.
机译:人类脐带血(HUCB)是成人干细胞的极好来源,具有比骨髓干细胞年轻的优点。干细胞在损伤修复机制中的作用仍在研究中。我们评估了HUCB通过共培养干预成纤维细胞去分化可塑性的能力。直接和间接共培养分别维持24、48和72小时。MTT法检测共培养活力。提取信使RNA,用定量逆转录聚合酶链反应(qRT PCR)检测p16和p21基因的表达。直接或间接接触不影响成纤维细胞的活力。然而,这些直接和间接接触降低了p16和p21基因的表达。采用S形曲线对基因表达进行时间调节,并根据细胞培养类型建立基因表达的数学函数。这些结果表明,未成熟细胞(HUCB)通过直接接触或信号分子影响分化细胞,从而改变其行为和可塑性。因此,我们的数据可能有助于旁分泌效应,而不是众所周知的在干细胞治疗中负责损伤修复的效应。

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