Development of cell sheet constructs for layer by layer tissue engineering using the blood vessel as an experimental model 2

Chapter 4. Isolation and Characterisation Of Vascular Progenitors rapidly proliferated to confluence, and failed to express endothelial markers CD31 and CD144. 121 Chapter 4. Isolation and Characterisation Of Vascular Progenitors Figure 4-3: Primary culture of haemopoietic tissue in EGM. Following culture in endothelial cell media, outgrowth colonies were generated from (a) umbilical cord blood mononuclear cells with typical cobblestone morphology. Similarly, outgrowth cells could be generated from (b) fetal blood mononuclear cells, which appear to have a more heterogeneous phenotype, adopting a more spindle morphology. (c) When cultured in the same medium, fibroblastic cells quickly grew out from fetal liver mononuclear cells, and failed to express endothelial phenotype. 122 Chapter 4. Isolation and Characterisation Of Vascular Progenitors (a) (b) (c) (d) (e) (f) Figure 4-4: CFU assay Adherent colonies appeared after ten days of culture, and were stained with crystal violet to facilitate visualization. Cobblestone colonies were generated from cord blood (a, b). In contrast, outgrowth colonies from fetal blood (c) were heterogeneous in nature, adopting morphologies ranging from cobblestone, fibroblastic to spindle (d - f). 123 . cultured in the same medium, fibroblastic cells quickly grew out from fetal liver mononuclear cells, and failed to express endothelial phenotype. Chapter 4. Isolation and Characterisation Of Vascular. 4. Isolation and Characterisation Of Vascular Progenitors 122 Figure 4-3: Primary culture of haemopoietic tissue in EGM. Following culture in endothelial cell media, outgrowth colonies were. Isolation and Characterisation Of Vascular Progenitors 121 rapidly proliferated to confluence, and failed to express endothelial markers CD31 and CD144. Chapter 4. Isolation and Characterisation