■观察和监测细胞密度和形态的能力对于评估细胞培养物的健康和生产高质量是必不可少的。几十年的高产细胞培养。基于微载体的文化,用于大规模细胞扩增过程,与传统的基于可视化的方法不兼容,比如宽视场显微镜,由于它们的厚度和材料组成。
■这里,我们评估了商业聚苯乙烯微载体与定制明胶甲基丙烯酰基(gelMA)微载体的光学成像相容性,用于整个微载体表面的非破坏性和非侵入性可视化,直接单元格枚举,和间充质干细胞/基质细胞的亚细胞可视化。
■进行了聚苯乙烯和gelMA微载体的Mie散射和波前误差模拟,以评估基于弹性散射的粘附细胞成像的潜力。蔡司Z.1光片显微镜适用于使用平面侧面照明的无标签弹性散射对比度进行光片断层扫描,以实现光学切片,并允许非侵入性和非破坏性,在托托,三维,在微载体上培养的细胞的高分辨率可视化。
■聚苯乙烯微载体使用荧光或弹性散射对比防止微载体远端一半上的细胞可视化,而gelMA微载体允许使用光片荧光显微镜和断层扫描对细胞形态进行高保真可视化和细胞密度定量。
■光学质量的gelMA微载体和无标签光片层析成像的组合将有助于增强对生物反应器-微载体细胞培养过程的控制。
UNASSIGNED: The ability to observe and monitor cell density and morphology has been imperative for assessing the health of a cell culture and for producing high quality, high yield cell cultures for decades. Microcarrier-based cultures, used for large-scale cellular expansion processes, are not compatible with traditional visualization-based methods, such as widefield microscopy, due to their thickness and material composition.
UNASSIGNED: Here, we assess the optical imaging compatibilities of commercial polystyrene
microcarriers versus custom-fabricated gelatin methacryloyl (gelMA)
microcarriers for non-destructive and non-invasive visualization of the entire microcarrier surface, direct cell enumeration, and sub-cellular visualization of mesenchymal stem/stromal cells.
UNASSIGNED: Mie scattering and wavefront error simulations of the polystyrene and gelMA
microcarriers were performed to assess the potential for elastic scattering-based imaging of adherent cells. A Zeiss Z.1 light-sheet microscope was adapted to perform light-sheet tomography using label-free elastic scattering contrast from planar side illumination to achieve optical sectioning and permit non-invasive and non-destructive, in toto, three-dimensional, high-resolution visualization of cells cultured on
microcarriers.
UNASSIGNED: The polystyrene microcarrier prevents visualization of cells on the distal half of the microcarrier using either fluorescence or elastic scattering contrast, whereas the gelMA microcarrier allows for high fidelity visualization of cell morphology and quantification of cell density using light-sheet fluorescence microscopy and tomography.
UNASSIGNED: The combination of optical-quality gelMA
microcarriers and label-free light-sheet tomography will facilitate enhanced control of bioreactor-microcarrier cell culture processes.