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
121

rapidly proliferated to confluence, and failed to express endothelial markers CD31
and CD144.
Chapter 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 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.
Chapter 4. Isolation and Characterisation Of Vascular Progenitors
123



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).
(a)

(
b
)

(
c
)

(
d
)

(
e
)

(f
)