Krešić Nina, Šimić Ivana, Lojkić Ivana, Bedeković Tomislav
Virology Department, Croatian Veterinary Institute, Savska Cesta 143, 10,000 Zagreb, Croatia.
Stem Cells Int. 2017;2017:4176292. doi: 10.1155/2017/4176292. Epub 2017 Jan 26.
Although canine adipose derived stem cells (cASCs) morphology characteristics and differentiation ability are well documented, transcriptome alterations of undifferentiated cASCs during ex vivo cultivation remain unknown. Here we demonstrate, for the first time, the transcriptome composition of isolated cASCs in undifferentiated state originating from six donors. Transcriptome changes were monitored during ex vivo cultivation between passage 3 (P3) and P5, which are mostly used in therapy. Influence of donors' age in given passage number on transcriptome composition was also investigated. Cultivation from P3 to P5 resulted in 16 differentially expressed genes with cooverexpression of pluripotency and self-renewal transcription factors genes SOX2 and POU5F1 dominant in old donors' cells. Furthermore, cASCs demonstrated upregulation of IL-6 in young and old donors' cells. In addition, ex vivo cultivation of cASCs revealed well-known morphological alterations accompanied with decrease in expression of CD90 and CD44 markers in P4 and higher monitored by flow cytometry and successful osteo- and chondrodifferentiation but inefficient adipodifferentiation in P3. Our results revealed the impact of ex vivo cultivation on nature of cells. Correlation of transcriptome changes with secretome composition is needed and its further impact on therapeutic potential of cASCs remains to be evaluated in clinical trials.
尽管犬脂肪来源干细胞(cASCs)的形态特征和分化能力已有充分记载,但体外培养过程中未分化cASCs的转录组变化仍不清楚。在此,我们首次展示了来自六个供体的未分化状态下分离的cASCs的转录组组成。在体外培养过程中监测了第3代(P3)和第5代(P5)之间的转录组变化,这两代细胞在治疗中最常使用。还研究了给定传代次数下供体年龄对转录组组成的影响。从P3培养到P5导致16个差异表达基因,多能性和自我更新转录因子基因SOX2和POU5F1的共过表达在老年供体细胞中占主导地位。此外,cASCs在年轻和老年供体细胞中均表现出IL-6的上调。另外,cASCs的体外培养显示出众所周知的形态学改变,伴随着第4代及更高代中CD90和CD44标志物表达的降低,通过流式细胞术监测,并且在P3中成功进行了成骨和成软骨分化,但脂肪分化效率低下。我们的结果揭示了体外培养对细胞性质的影响。需要转录组变化与分泌组组成的相关性,并且其对cASCs治疗潜力的进一步影响仍有待在临床试验中评估。