Transplantation of Genetically Engineered Cardiac Fibroblasts Producing Recombinant Human Erythropoietin to Repair the Infarcted Myocardium - Genetic Engineering: Recent Developments in Applications - page 197

Figure 3. Four weeks after cell transplantation, rhEPO expression promotes neo-angiogenesis in vivo. (A)
α -Smooth muscle actin staining (brown) in infarcted hearts (infarct border zones). Arrows show stained
blood vessels. (a) Saline injection; (b) non-transduced cell injection; (c) rhEPO-transduced cell injection; (d)
quantitative analysis. (B) Lectin (BS-1) staining (brown) in infarcted hearts (infarct border zones). Arrows show
stained blood vessels. (a) Saline injection; (b) non-transduced cell injection; (c) rhEPO-transduced cell injection;
(d) quantitative analysis.
Apoptosis in infarcted hearts was detected by active caspase-3 staining with
polyclonal antibodies detecting only the active (effector) form of caspase-3 (Fig-
ure 4A) [25,26]. Human tonsil tissue was used as a positive control (Figure 4A,
a). RhEPO-transduced cell transplantation resulted in a significant decrease in the
number of active-caspase-3 positive cells (24 ± 6 versus 176 ± 20 versus 110 ± 27
cells per mm2 in saline and non-transduced cell injections, p < 0.05; Figure 4C,
a). The staining revealed mainly nuclear localization of active caspase-3 (Figure
4A) [26]. Apoptosis was also evaluated by commercially available TUNEL kit
(Figure 4B). Rat mammary gland tissue served as a positive control (Figure 4B,
a). Similar to active caspase-3 staining, TUNEL staining showed that rhEPO-
transduced cell transplantation resulted in a significant decrease in the number of