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FACS-purified myoblasts producing controlled VEGF levels induce safe and stable angiogenesis in chronic hind limb ischemia.

Wolff T, Mujagic E, Gianni-Barrera R, Fueglistaler P, Helmrich U, Misteli H, Gurke L, Heberer M, Banfi A - J. Cell. Mol. Med. (2012)

Bottom Line: Vessel density was doubled in engrafted areas, but only approximately 0.1% of muscle volume showed cell engraftment, explaining why no increase in total blood flow was observed.In conclusion, the use of FACS-purified myoblasts granted the cell-by-cell control of VEGF expression levels, which ensured long-term safety in a model of chronic ischemia.Based on these results, the total number of implanted cells required to achieve efficacy will need to be determined before a clinical application.

View Article: PubMed Central - PubMed

Affiliation: Cell and Gene Therapy, Department of Biomedicine, Basel University Hospital, Basel, Switzerland.

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Myoblast engraftment is minimal compared to muscle size. Myoblast engraftment was revealed by X-Gal staining (blue) on frozen sections of the same medial hamstring muscles shown in Figure 4. (A) A low magnification image of a cross-section of the entire muscle implanted with transduced myoblasts shows cell engraftment only along the needle tracts of cell injections (white arrows). Higher magnification images showing similarly low engraftment rates with control cells (B), reference clone cells (C), the primary transduced population (D) and purified cells (E). Size bars 4 mm (A) and 400 μm (B–E). (F) Engraftment was quantified on all X-Gal stained sections and expressed as a percent of the total muscle volume (n = 3–5).
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fig06: Myoblast engraftment is minimal compared to muscle size. Myoblast engraftment was revealed by X-Gal staining (blue) on frozen sections of the same medial hamstring muscles shown in Figure 4. (A) A low magnification image of a cross-section of the entire muscle implanted with transduced myoblasts shows cell engraftment only along the needle tracts of cell injections (white arrows). Higher magnification images showing similarly low engraftment rates with control cells (B), reference clone cells (C), the primary transduced population (D) and purified cells (E). Size bars 4 mm (A) and 400 μm (B–E). (F) Engraftment was quantified on all X-Gal stained sections and expressed as a percent of the total muscle volume (n = 3–5).

Mentions: In order to investigate why the angiogenic induction observed by histological analysis was not accompanied by an increase in muscle blood flow, we examined the extent of stable engraftment of transduced myoblasts in the medial hamstring muscles 3 months after injection (n = 4 per condition). As can be seen in Figure 6A, X-Gal+ muscle fibres could only be found along the needle tract of the intramuscular injections (white arrows). The engraftment pattern was similar for the control rCD8 cells, the reference clone, the primary rVICD8 population and the purified population (Fig. 6B–E, respectively) and in all cases accounted for only approximately 0.1–0.2% of the total volume of the treated muscle (Fig. 6F).


FACS-purified myoblasts producing controlled VEGF levels induce safe and stable angiogenesis in chronic hind limb ischemia.

Wolff T, Mujagic E, Gianni-Barrera R, Fueglistaler P, Helmrich U, Misteli H, Gurke L, Heberer M, Banfi A - J. Cell. Mol. Med. (2012)

Myoblast engraftment is minimal compared to muscle size. Myoblast engraftment was revealed by X-Gal staining (blue) on frozen sections of the same medial hamstring muscles shown in Figure 4. (A) A low magnification image of a cross-section of the entire muscle implanted with transduced myoblasts shows cell engraftment only along the needle tracts of cell injections (white arrows). Higher magnification images showing similarly low engraftment rates with control cells (B), reference clone cells (C), the primary transduced population (D) and purified cells (E). Size bars 4 mm (A) and 400 μm (B–E). (F) Engraftment was quantified on all X-Gal stained sections and expressed as a percent of the total muscle volume (n = 3–5).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3823097&req=5

fig06: Myoblast engraftment is minimal compared to muscle size. Myoblast engraftment was revealed by X-Gal staining (blue) on frozen sections of the same medial hamstring muscles shown in Figure 4. (A) A low magnification image of a cross-section of the entire muscle implanted with transduced myoblasts shows cell engraftment only along the needle tracts of cell injections (white arrows). Higher magnification images showing similarly low engraftment rates with control cells (B), reference clone cells (C), the primary transduced population (D) and purified cells (E). Size bars 4 mm (A) and 400 μm (B–E). (F) Engraftment was quantified on all X-Gal stained sections and expressed as a percent of the total muscle volume (n = 3–5).
Mentions: In order to investigate why the angiogenic induction observed by histological analysis was not accompanied by an increase in muscle blood flow, we examined the extent of stable engraftment of transduced myoblasts in the medial hamstring muscles 3 months after injection (n = 4 per condition). As can be seen in Figure 6A, X-Gal+ muscle fibres could only be found along the needle tract of the intramuscular injections (white arrows). The engraftment pattern was similar for the control rCD8 cells, the reference clone, the primary rVICD8 population and the purified population (Fig. 6B–E, respectively) and in all cases accounted for only approximately 0.1–0.2% of the total volume of the treated muscle (Fig. 6F).

Bottom Line: Vessel density was doubled in engrafted areas, but only approximately 0.1% of muscle volume showed cell engraftment, explaining why no increase in total blood flow was observed.In conclusion, the use of FACS-purified myoblasts granted the cell-by-cell control of VEGF expression levels, which ensured long-term safety in a model of chronic ischemia.Based on these results, the total number of implanted cells required to achieve efficacy will need to be determined before a clinical application.

View Article: PubMed Central - PubMed

Affiliation: Cell and Gene Therapy, Department of Biomedicine, Basel University Hospital, Basel, Switzerland.

Show MeSH
Related in: MedlinePlus