人绒毛膜绒毛和羊水间充质干细胞在体外大量扩增后不易发生转化。
Human mesenchymal stem cells from chorionic villi and amniotic fluid are not susceptible to transformation after extensive in vitro expansion.
机构信息
Clinica di Ematologia, Dipartimento Scienze Mediche e Chirurgiche, Università Politecnica delle Marche, Ancona, Italy.
出版信息
Cell Transplant. 2011;20(5):643-54. doi: 10.3727/096368910X536518. Epub 2010 Nov 5.
Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering. Increasing evidence suggests that MSCs isolated from fetal tissues are more plastic and grow faster than adult MSCs. In this study, we characterized human mesenchymal progenitor cells from chorionic villi (CV) and amniotic fluid (AF) isolated during the first and second trimesters, respectively, and compared them with adult bone marrow-derived MSCs (BM). We evaluated 10 CV, 10 AF, and 6 BM samples expanded until the MSCs reached senescence. We used discarded cells from prenatal analyses for all the experiments. To evaluate the replicative stability of these cells, we studied the telomerase activity, hTERT gene transcription, and telomere length in these cells. Spontaneous chromosomal alterations were excluded by cytogenetic analysis. We studied the expression of c-myc and p53, tumor-associated genes, at different passage in culture and the capacity of these cells to grow in an anchorage-independent manner by using soft agar assay. We isolated homogeneous populations of spindle-shaped CV, AF, and BM cells expressing mesenchymal immunophenotypic markers throughout the period of expansion. CV cells achieved 14 ± 0.9 logs of expansion in 118 days and AF cells achieved 21 ± 0.9 logs in 118 days, while BM cells achieved 11 × 0.4 logs in 84 days. Despite their high proliferation capacity, fetal MSCs showed no telomerase activity, no hTERT and c-myc transcriptions, and maintained long, stable telomeres. A constant expression level of p53 and a normal karyotype were preserved throughout long-term expansion, suggesting the safety of fetal MSCs. In conclusion, our results indicate that fetal MSCs could be an alternative, more accessible resource for cell therapy and regenerative medicine.
间充质干细胞(MSCs)是细胞治疗和组织工程的有前途的候选者。越来越多的证据表明,从胎儿组织中分离的 MSCs 比成人 MSCs 更具可塑性且生长更快。在这项研究中,我们分别对来自第一和第二孕期的绒毛膜绒毛(CV)和羊水(AF)的人间质祖细胞进行了特征描述,并将其与成人骨髓来源的 MSCs(BM)进行了比较。我们评估了 10 个 CV、10 个 AF 和 6 个 BM 样本,直到 MSCs 达到衰老为止。我们使用产前分析中丢弃的细胞进行了所有实验。为了评估这些细胞的复制稳定性,我们研究了这些细胞中的端粒酶活性、hTERT 基因转录和端粒长度。通过细胞遗传学分析排除了自发的染色体改变。我们研究了在不同培养传代时这些细胞中肿瘤相关基因 c-myc 和 p53 的表达,以及这些细胞在无锚定生长方式下的生长能力,使用软琼脂测定法。我们分离了表达间充质免疫表型标记的均一群体的 CV、AF 和 BM 细胞,这些细胞在整个扩增过程中都得到了扩增。CV 细胞在 118 天内实现了 14 ± 0.9 对数的扩增,AF 细胞在 118 天内实现了 21 ± 0.9 对数的扩增,而 BM 细胞在 84 天内实现了 11 × 0.4 对数的扩增。尽管具有高增殖能力,但胎儿 MSCs 没有端粒酶活性,没有 hTERT 和 c-myc 转录,并且保持了长而稳定的端粒。在长期扩增过程中,p53 的表达水平保持恒定且核型正常,提示胎儿 MSCs 的安全性。总之,我们的结果表明,胎儿 MSCs 可能是细胞治疗和再生医学的一种替代、更易获得的资源。