Tumour cells, with or without TMV, were prepared as
above, washed and resuspended in 100 μl of PBS. Then,
cells were mixed with 400 μl of Matrigel Matrix (BD Bio-sciences) and injected subcutaneously into the abdominal
middle line of NOD SCID mice. After 6 days mice were
euthanized, Matrigel Matrix pellets were cut out and then
400 μl of Cell Recovery Solution (BD Biosciences) was
added and left at 4 °C for 7 days. Then, Matrigel Matrix
implants were suspended in Drabkin’s solution (Sigma-Aldrich) for 30 min, after that spun down and the con-tent of haemoglobin was determined by measuring the
absorbance (O.D.) at 540 nm in U-1800 spectrophotom-eter (Hitachi, Tokyo, Japan) [20]. Independently Matrigel
plugs after cutting out were photographed and then fixed
in formalin, paraffin embedded, serial 3 μm sections were
cut and stained with hematoxylin and eosin