University of Twente Student Theses


Analysing the Intracellular Crowding of Primary Chondrocytes from Passage 2 to Passage 8 using Holotomography

Elderhorst, Anouk (2023) Analysing the Intracellular Crowding of Primary Chondrocytes from Passage 2 to Passage 8 using Holotomography.

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Abstract:It is known that primary chondrocytes dedifferentiate over time with increasing cell passage, in in vitro monolayer cell culture . Dedifferentiation is characterized by a loss of cell phenotype, including a change in extracellula r matrix production, cell morphology and the cells increasing inability to respond to chondrogenic differentiation medium. While dedifferentiating chondrocytes show an increase in cell surface area, little is known about their cell volume and intracellular molecular crowding. This bachelor assignment focussed on analysing the cell volume and dry mass over time of human primary chondrocytes of passage 2, 4, 6 and 8 , to investigate if there is a correlation between chondrocyte dedifferentiation a nd changes in intracellular crowding. It was found that between P2 and P8 there was an overall decrease in intracellular crowding resulting from an ove r all de crease in cell volume and drymass . For the analysis the new holotomography microscope, called T omocube, was used. As part of the reasearch, the sensitivity of the holotomography microscope was tested, to optimize the protocol for imaging. It was shown that from ana lysis with the Tomocube mi croscope it is possible to show differences in cell volume within a time sequence of 5 minute s Also, the analysis software of the Tomocube was compared to Imaris, a commonly used imag ing analysi s software. The cell volume obtained with the Tomocube analysis software was similar to the cell volume obtained after surface reconstruction in Imaris. Finally, the attachment and spreading of primary chondrocytes of P2 hPC s were observed with a 32 hour long time lapse. Between one and eight hours there is hardly any fluctation in cell volume observed, fluctations started after eight hours.
Item Type:Essay (Bachelor)
Faculty:TNW: Science and Technology
Programme:Biomedical Technology BSc (56226)
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