Autotransplantation of the adipose tissue derived stem cells for chronic

wounds healing.

L. Masłowski 1, M.Paprocka2, W. Witkiewicz1, D. Duś2, R. Grendziak1, J. Kubiak1, A. Buczyńska1, E. Wojtowicz-Prus1, A. Czarnecka1 1Regional Specialist Hospital, Research and Development Centre in Wroclaw, 2Ludwik Hirszfeld Institute of Immunology and Experimental Therapy of Polish Academy of Science WROVASC Integrated Center of Cardiovascular Medicine

Mesenchymal stem cells have now been isolated from various tissues including: - bone marrow, - peripheral blood, - cord blood, - muscle, - skin, - synovium - adipose tissue

Adipose tissue could be one of the most suitable cell sources for cell therapy, because of its - easy accessibility, - minimal morbidity and - abundance of mesenchymal stem cells.

The study group consisted of 21 patients: • 14 men and 7 women; • mean age 62,9 + 8,4y; • with chronic wounds of vascular origin: • 13 of the diabetic foot and 8 of venous ulcers. After autologous cells transplantation all patients were 6 months of follow-up and received standard local and general treatment.

• Adipose tissue was harvested by aspiration after infiltration by local anesthetic and tumescend isotonic solution.

• Stem and regenerative cells were separated using closed and fully automated system CELLUTION 800 from Cytori Therapeutics USA.

• Cell suspension concentrate volume of 5 ml was implanted to subcutaneous tissue around the wound and the wound bed. The phenotype of the cells obtained was determined immediately after separation and after 7 days of culture on RPMI with 10% fetal calf serum.

Clinical results: In patients responding to the regenerative therapy healing acceleration were

obserwed to 10-12 weeks after autotransplantation.

• 75% of patients showed improvement of venous ulcers (6 of 8)

including 1 completely healed and 5 with >50% reduction of ulcer area. 1 patient did not improve progression was observed in one of them.

•in patients with diabetic foot improvement was observed in 76,9% including 4 completely healed and 6 with >50% reduction of ulcer

area. 2 patients there was no improvement and 1 presented progreession of necrotic chandes resulting in amputation.

Laboratory results: The phenotype of the cells after separation: The phenotype of the cells after culture:

CD45

CD31

CD34

CD133

CD146

CD105

CD90

śr + SD 24,82 + 5,86 22,27 +

10,02

65,55 +

24,05

0,27 + 0,65 29,73 +

23,74

33,36 +

25,15

68,43 +

22,66

CD45

CD31

CD34

CD133

CD146

CD105

CD90

śr + SD 9,5 + 10,4 9,1 + 11,3 38,3 + 28,4 0,0 + 0,0 16,5 + 8,9 81,8 + 31,0 93,5 + 8,3

Autotransplantation of adipose tissue stem cells is a promising treatment for chronic wounds,

however fragile the effect of a single application of stem cells prompted us to search for methods of

improvement. The use of cells from the culture seems to be a good continuation of the current clinical study.