Activated mesenchymal stem cells for treating limb ischemia
A method for treating limb ischemia in a patient in need of such treatment by administering metamizole treated mesenchymal stem cells to the patient. The method provides a pharmaceutical product for angiogenesis growth to replace occluded blood vessels for avoiding limb amputation. Secretion of angiogenic growth factors VEGFA, HGF, bFGF, TEK are stimulated and the levels of pro-inflammatory cytokines IF6, CXCF8, CCF2, IL1-RN are reduced by activation of mesenchymal stem cells (MSCs). According to treatment results, MSCs produce proteins and signaling molecules for new blood vessel growth that accelerate the growth of new arteries.
1 . A method for promoting angiogenesis in a limb of a human having restricted blood supply, the method comprising:
(i) treating human adipose-derived mesenchymal stem cells (AdMSCs) with lipopolysaccharide (LPS) to heighten levels of inflammatory cytokines in the AdMSCs thereby producing LPS treated AdMSCs,
(ii) treating the LPS treated AdMSCs with metamizole to suppress expression of pro-inflammatory cytokines CCL2, CCL3, TNF, IL6, IL1A, and IL1B and thereby producing metamizole treated AdMSCs, and
(iii) intramuscularly administering to the limb of the human the metamizole treated AdMSCs in an amount effective to stimulate formation of new blood vessels in the limb of the human.
2 . The methods according to claim 1 , wherein the metamizole treated AdMSCs are injected into a muscle of the subject in a dose of 1 million cells per kg of the subject.
3 . The method according to claim 1 , wherein the metamizole treated AdMSCs are administered to the subject in an amount of 0.75-1.5 million cells per kg of body weight of the subject.
4 . The method according to claim 3 , wherein the AdMSCs are produced by a process comprising the following steps:
(i) collecting fat tissue
(ii) separating mesenchymal stem cells (MSCs) from the fat tissue to produce adipose-derived mesenchymal stem cells (AdMSCs), and
(iii) expanding the AdMSCs.
5 . The method according to claim 1 , wherein in step (i) the AdMSCs are stimulated with 0.1 μg/ml of the LPS for 2 hours.
6 . The method according to claim 5 , wherein in step (ii) the LPS treated AdMSCs are contacted with 10 μM of the metamizole for 24 hours.