Raposio E, Guida C, Coradeghini R, Scanarotti C, Parodi A, Baldelli I, Fiocca R, Santi P L
Tissue Engineering Laboratory, Plastic Surgery Division, DICMI, University of Genoa, Genoa, Italy.
Cell Prolif. 2008 Oct;41(5):739-54. doi: 10.1111/j.1365-2184.2008.00547.x. Epub 2008 Jul 31.
Adipose tissue is the most abundant and accessible source of adult stem cells. Human processed lipoaspirate contains pre-adipocytes that possess one of the a characteristic pathways of multipotent adult stem cells and are able to differentiate in vitro into mesenchymal and also neurogenic lineages. Because stem cells have great potential for use in tissue repair and regeneration, it would be significant to be able to obtain large amounts of these cells in vitro. As demonstrated previously, purine nucleosides and nucleotides mixtures can act as mitogens for several cell types. The aim of this study was to evaluate the effects of polydeoxyribonucleotides (PDRN), at appropriate concentrations, on human pre-adipocytes grown in a controlled medium, also using different passages, so as to investigate the relationship between the effect of this compound and cellular senescence, which is the phenomenon when normal diploid cells lose the ability to divide further.
Human pre-adipocytes were obtained by liposuction. Cells from different culture passages (P6 and P16) were treated with PDRN at different experimental times. Cell number was evaluated for each sample by direct counting after trypan blue treatment. DNA assay and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test were also carried out in all cases.
PDRN seemed to promote proliferation of human pre-adipocytes at both passages, but cell population growth increased in pre-adipocyte at P16, after 9 days as compared to control. Our data suggest that PDRN could act as a pre-adipocyte growth stimulator.
脂肪组织是成体干细胞最丰富且易于获取的来源。人加工过的抽脂物含有前脂肪细胞,这些前脂肪细胞具有多能成体干细胞的一种特征性途径,并且能够在体外分化为间充质谱系以及神经谱系。由于干细胞在组织修复和再生方面具有巨大的应用潜力,因此能够在体外大量获取这些细胞具有重要意义。如先前所示,嘌呤核苷和核苷酸混合物可作为多种细胞类型的促细胞分裂剂。本研究的目的是评估适当浓度的聚脱氧核糖核苷酸(PDRN)对在受控培养基中生长的人前脂肪细胞的影响,同时使用不同传代的细胞,以研究该化合物的作用与细胞衰老之间的关系,细胞衰老是指正常二倍体细胞失去进一步分裂能力的现象。
通过抽脂获取人前脂肪细胞。来自不同培养传代(P6和P16)的细胞在不同实验时间用PDRN处理。经台盼蓝处理后,通过直接计数评估每个样品的细胞数量。在所有情况下还进行了DNA检测和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐试验。
PDRN似乎在两个传代水平上均促进人前脂肪细胞的增殖,但与对照组相比,P16的前脂肪细胞在9天后细胞群体生长增加。我们的数据表明,PDRN可作为前脂肪细胞生长刺激剂。