The effects of allogenic stem cells in a murine model of hind limb diabetic ischemic tissue

Jesús Álvarez García, Soledad García Gómez-Heras, Luis Riera Del Moral, Carlota Largo, Damián García-Olmo, Mariano García-Arranz, Jesús Álvarez García, Soledad García Gómez-Heras, Luis Riera Del Moral, Carlota Largo, Damián García-Olmo, Mariano García-Arranz

Abstract

Background: Diabetes is one of the major risk factors for peripheral arterial disease. In patients in whom surgery cannot be performed, cell therapy may be an alternative treatment. Since time is crucial for these patients, we propose the use of allogenic mesenchymal cells.

Methods: We obtained mesenchymal cells derived from the fat tissue of a healthy Sprague-Dawley rat. Previous diabetic induction with streptozotocin in 40 male Sprague-Dawley rats, ligation plus left iliac and femoral artery sections were performed as a previously described model of ischemia. After 10 days of follow-up, macroscopic and histo-pathological analysis was performed to evaluate angiogenic and inflammatory parameters in the repair of the injured limb. All samples were evaluated by the same blind researcher. Statistical analysis was performed using the SPSS v.11.5 program (P < 0.05).

Results: Seventy percent of the rats treated with streptozotocin met the criteria for diabetes. Macroscopically, cell-treated rats presented better general and lower ischemic clinical status, and histologically, a better trend towards angiogenesis, greater infiltration of type 2 macrophages and a shortening of the inflammatory process. However, only the inflammatory variables were statistically significant. No immunological reaction was observed with the use of allogeneic cells.

Discussion: The application of allogeneic ASCs in a hind limb ischemic model in diabetic animals shows no rejection reactions and a reduction in inflammatory parameters in favor of better repair of damaged tissue. These results are consistent with other lines of research in allogeneic cell therapy. This approach might be a safe, effective treatment option that makes it feasible to avoid the time involved in the process of isolation, expansion and production of the use of autologous cells.

Keywords: Allogeneic stem cells; Limb ischemia.

Conflict of interest statement

Prof. D. García Olmo and Dr. M. García Arranz have applied for two patents related to Adipose Derived Mesenchymal Stem Cells titled “Identification and Isolation of Multipotent Cells from Non-Osteochondral Mesenchymal Tissue” (WO 2006/057649) and “Use of Adipose Tissue-Derived Stromal Stem Cells in Treating Fistula” (WO 2006/136244). The remaining authors have no other financial or competing interests to declare.

Figures

Figure 1. Images of the surgical procedure.
Figure 1. Images of the surgical procedure.
Figure 2. Flow chart.
Figure 2. Flow chart.
ASC, adipose derived stem cells.
Figure 3. Clinical evolution.
Figure 3. Clinical evolution.
(A) Physical parameters; (B) ischemic parameters.
Figure 4. Macroscopic appearance of the treated…
Figure 4. Macroscopic appearance of the treated limb.
(A) 10 Days after surgery without cellular treatment; (B) 10 Days after surgery and treatment with cells.
Figure 5. Histopathological results.
Figure 5. Histopathological results.
(A) Treatment group. Low number of inflammatory cells (arrow), capillaries and and neoformed arterioles (asterisk). Hematoxiline-eosine staining, 20×. (B) Control Group l, greater number of inflammatory infiltrate; inflammatory cells (arrow), capillaries and neoformed arterioles (asterisk). Hematoxiline-eosine staining, 20×. (C–D) Expression of macrophagues M1/CD68+ in the larger photographs and M2/CD206+ expression in the small-right photographs. Cicatricial area of treated group (C) and control group (D), 20×. (E–F) Regional lymph nodes of treated group (E) and control group (F), 20×.

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