Coupland Paul G, Fisher Karen A, Jones D Rhodri E, Aylott Jonathan W
Laboratory of Biophysics and Surface Analysis, School of Pharmacy, University of Nottingham, University Park, Nottingham NG72RD, UK.
J Control Release. 2008 Sep 10;130(2):115-20. doi: 10.1016/j.jconrel.2008.06.018. Epub 2008 Jun 27.
The aim of this study was to demonstrate that flow cytometry and confocal microscopy could be applied in a complementary manner to analyse the internalisation of polymeric nanosensors in mesenchymal stem cells (MSC). The two techniques are able to provide en masse data analysis of nanosensors from large cell populations and detailed images of intracellular nanosensor localisation, respectively. The polyacrylamide nanosensors used in this investigation had been modified to contain free amine groups which were subsequently conjugated to Tat peptide, which acted as a delivery vector for nanosensor internalisation. Flow cytometry was used to confirm the health of MSC culture and assess the impact of nanosensor internalisation. MSC were characterised using fluorescently tagged CD cell surface markers that were also used to show that nanosensor internalisation did not negatively impact on MSC culture. Additionally it was shown that flow cytometry can be used to measure fluorophores located both on the cell surface and internalised within the cell. Complementary data was obtained using confocal microscopy to confirm nanosensor internalisation within MSC.
本研究的目的是证明流式细胞术和共聚焦显微镜可以以互补的方式应用于分析聚合物纳米传感器在间充质干细胞（MSC）中的内化情况。这两种技术分别能够对来自大量细胞群体的纳米传感器进行整体数据分析以及提供细胞内纳米传感器定位的详细图像。本研究中使用的聚丙烯酰胺纳米传感器经过修饰，含有游离胺基，随后与Tat肽缀合，Tat肽作为纳米传感器内化的递送载体。流式细胞术用于确认MSC培养的健康状况并评估纳米传感器内化的影响。使用荧光标记的CD细胞表面标志物对MSC进行表征，这些标志物也用于表明纳米传感器内化不会对MSC培养产生负面影响。此外，还表明流式细胞术可用于测量位于细胞表面和细胞内内化的荧光团。使用共聚焦显微镜获得互补数据，以确认纳米传感器在MSC内的内化情况。
