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Results 32 Further

Results 32 Further parameters for describing the cells shape (cell area, circularity, ferret’s diameter and aspect ratio) from all passages in both settings were acquired and are provided as supplementary data (Supp.Tab. 2 & Supp.Fig. 1, page 56 & 57). Tab. 4: Quantification of cell area and aspect ratio of 100 hMSCs per quarter in both culture conditions. Mean & SD. Time period Cell area (x10 4 µm 2 ) Aspect ratio Normoxia Hypoxia Normoxia Hypoxia 1. Quarter 0.609 ± 0.419 0.440 ± 0.212 3.636 ± 1.775 3.819 ± 1.801 2. Quarter 0.823 ± 0.466 0.554 ± 0.238 4.095 ± 2.472 3.953 ± 2.469 3. Quarter 1.392 ± 0.804 0.598 ± 0.368 2.487 ± 1.227 3.111 ± 1.640 4. Quarter 1.344 ± 0.967 1.419 ± 0.654 3.750 ± 2.320 2.575 ± 1.495 3.2.4. Cell volume measurement Since cells are three-dimensional objects and thus cell volume describes the last morphological parameter. Therefore, 25 cells were measured for cell volume and cell height on different ECM proteins (collagen I, fibronectin and laminin) with white-light confocal microscopy. Normal ionized cell culture plastic served as a control. Exemplary topographical images of normoxic and hypoxic cultured cells on different ECM coated slides are shown in figure 16. Cell heights were continuously lower in hypoxia compared to normoxic cultured cells, whereas only laminin coating revealed a significant difference (p=0.0254). Table 5 shows all mean cell height values. Tab. 5: Cell heights of normoxic and hypoxic cultured hMSCs on different surfaces. Cell height [µm] Parameters NUNC Collagen I Fibronectin Laminin N H N H N H N H Mean 1.27 1.08 1.38 1.11 1.20 1.06 1.52 1.21 Median 1.21 1.03 1.20 1.00 1.20 1.02 1.44 1.06 SD 0.45 0.36 0.65 0.58 0.37 0.22 0.47 0.47

Results 33 Fig. 16: Exemplary white light confocal images of normoxic or hypoxic cultured hMSCs on different substrates. Overall, normoxic cultured cells were higher on polystyrene surfaces (NUNC) and collagen I, fibronectin or laminins coated glass slides. The colour bar represents the cells height. Bar: 50 µm.

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