Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, Cheras 56000, Kuala Lumpur, Malaysia.
Tissue Cell. 2012 Jun;44(3):156-63. doi: 10.1016/j.tice.2012.02.001. Epub 2012 Mar 7.
Tracking of transplanted cells has become an important procedure in cell therapy. We studied the in vitro dye retention, survival and in vivo tracking of stem cells with PKH26 dye. Sheep BMSCs and ADSCs were labeled with 2, 4 and 8 μmol of PKH26 and monitored for six passages. Labeled BMSCs and ADSCs acquired mean cumulative population doubling of 12.7±0.4 and 14.6±0.5; unlabeled samples had 13.8±0.5 and 15.4±0.6 respectively. Upon staining with 2, 4 and 8 μmol PKH26, BMSCs had retentions of 40.0±5.8, 60.0±2.9 and 95.0±2.9%, while ADSCs had 92.0±1.2, 95.0±1.2 and 98.0±1.2%. ADSCs retentions were significantly higher at 2 and 4 μmol. On dye retention comparison at 8 μmol and 4 μmol for BMSCs and ADSCs; ADSCs were significantly higher at passages 2 and 3. The viability of BMSCs reduced from 94.0±1.2% to 90.0±0.6% and ADSCs from 94.0±1.2% to 52.0±1.2% (p<0.05) after 24h. BMSCs had significant up regulation of the cartilage genes for both the labeled and the unlabeled samples compared to ADSCs (p<0.05). PKH26 fluorescence was detected on the resected portions of the regenerated neo-cartilage. The recommended concentration of PKH26 for ADSCs is 2 μmol and BMSCs is 8 μmol, and they can be tracked up to 49 days.
细胞示踪已成为细胞治疗中的一个重要程序。我们研究了 PKH26 染料对干细胞的体外染料保留、存活和体内示踪。绵羊骨髓间充质干细胞和脂肪来源间充质干细胞用 2、4 和 8 μmol 的 PKH26 进行标记，并监测了 6 个传代。标记的骨髓间充质干细胞和脂肪来源间充质干细胞的平均累积群体倍增分别为 12.7±0.4 和 14.6±0.5；未标记的样本分别为 13.8±0.5 和 15.4±0.6。用 2、4 和 8 μmol PKH26 染色后，骨髓间充质干细胞的保留率分别为 40.0±5.8、60.0±2.9 和 95.0±2.9%，而脂肪来源间充质干细胞分别为 92.0±1.2、95.0±1.2 和 98.0±1.2%。在 2 和 4 μmol 时，脂肪来源间充质干细胞的保留率明显更高。在骨髓间充质干细胞和脂肪来源间充质干细胞的 8 μmol 和 4 μmol 染料保留比较中，脂肪来源间充质干细胞在第 2 和第 3 代的保留率明显更高。骨髓间充质干细胞的活力从 94.0±1.2%降至 90.0±0.6%，脂肪来源间充质干细胞从 94.0±1.2%降至 52.0±1.2%（p<0.05），在 24 小时后。与脂肪来源间充质干细胞相比，标记和未标记的样本中骨髓间充质干细胞的软骨基因表达均显著上调（p<0.05）。在再生新软骨的切除部分检测到 PKH26 荧光。建议的 PKH26 浓度为脂肪来源间充质干细胞 2 μmol，骨髓间充质干细胞 8 μmol，可追踪至 49 天。
