Effects on immune cell viability, morphology and proliferation in a sub-microliter cell sampler system

• Main Author: Wiklund, Sofia
• Format: Others
• Language: English
• Published: Linköpings universitet, Institutionen för fysik, kemi och biologi 2013
• Subjects: Cell-picker; sub-microliter cell sampler system; immune cells; T cells; NK cells; B cells; cell viability; morphology; proliferation; surface coating; microchip; wells; silicon chip; single-cell analysis; flow system; fluorescence microscopy; cell staining; characterization
• Online Access: http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-89982

Today,   most traditional method used in the research of immune cells, such as flow   cytometry and microscopy, are based on average values of cell responses.   However, immune cells are heterogeneous and respond differently to a given   stimuli. There is also a risk that important, but rare, behaviors of   individual cells are missed when a larger population of immune cells is   analyzed. Also, flow cytometry and microscopy do not allow long-term survival   of cells; these methods lack the ability to do dynamic long-term analysis of   motile immune cells, i.e. studies of cell-cell interactions, morphology and proliferation.   In a   patient who is affected by cancer, the cell heterogeneity contributes to the   ability to battle various types of cancer or virus infections. In an   outbreak, immune cells recognize and kill tumor cells. However, the number of   specific immune cells is sometimes too few to kill all the tumor cells in a   successful way. One way to help these patients is to isolate, select out and   cultivate the active immune cells with capacity to kill tumor cells.   The   Cell Physic Laboratory (a part of the department of Applied Physics) at the   Royal Institute of Technology (KTH) has developed a method for single-cell   analysis where the immune cells are trapped in microwells in a silicon chip.   The immune cells are then studied by using fluorescence microscopy in an   inverted setup. The method enables high-throughput experiments due to the   parallelization. Furthermore, since the immune cells survive long periods in   the chip, the cells can be analyzed over several days up to weeks. The   research group has also developed a semi-automatic ‘cell-picker’. The   cell-picker will be used in combination with the developed method for   single-cell analysis, which enables picking of cells of interest. In this report, experiments for the characterization and evaluation of the biocompatibility of two generations of the cell-picker will be presented. The experiments include development of a protocol for the cell-picking process, studies of the survival time of transferred cells for both generation of the cell-picker and studies of surface coating in the chip in order to increase the biocompatibility. The preliminary results indicate that the cell-picker has potential to be used as a selection tool for immune cells of interest.