Centrifuged, filtered through a thin membrane washed in phosphate-buffered saline
Automated purification system-fitted with the single-use sterile kit having a rotor for cell washing. Then rinsing with human serum albumin-supplemented normal saline
Cells rinse and centrifuge
Washing with phosphate-buffered, centrifuged, and filtered
Culture medium
No
Mononuclear cell layer 1st. Cultured in alpha-modified essential medium containing 10% autologous serum 2nd. Cultured in alpha-MEM
Iscove’s modified Dulbecco’s medium (IMDM) with 10% fetal bovine serum, 10% horse serum, 5IM containing hydrocortisone, gentamycin-sulphate, and vancomycin in the medium
DMEM with 0.025% type 1 collagenase for 80 min at 37°C. Stromal vascular fraction cultured in DMEM to obtain enough ASCs. ASCs seeded into hydrogel matrix and cultured for expansion
DMEM with human serum 10%, streptomycin 1% solution, and stable glutamine 1% at 37°C under 5% CO2
Mode of administration and volume
Intramuscular injection, 1 ml into the ischemic limb, along post and anterior tibia
2 ml cells, injected in basal of foot ulcer and surrounding tissue
Intramuscular injection in M. gastrocnemius ipsilateral 1 ml each in deep
Allogeneic stem cell sheet put direct on the wound bed
Injected into dermoepidermal junction and homogenously the whole of the wound
Success rate
79% (29/37)
100% (41/41)
82% (18/22)
82% (18/22)
90% (9/10)
Reason for failure
Preexisting severe lymphopenia and thrombocytopenia Death due to coronary heart disease
No failure
Death (multiple organ failure due to sepsis postamputation)
Cellulitis, paresthesia, and uncontrolled diabetes cardiac arrest
Recurrent infections and necrosis leading to minor limb amputations
Adverse complications
Major limb amputation in patients with preexisting severe lymphopenia and thrombocytopenia
No acute or chronic adverse events related to BM-MSC or BM-NSC therapy
(Cellulitis, paresthesia, uncontrolled diabetes, and cardiac arrest, not related to treatment. Baseline clinical and lab changes not clinically meaningful) in both groups
No complications were observed in two years of follow ups
