Abstract Background Diabetic cutaneous ulcers are subjected to several physiological and biochemical defects, which contribute to wound chronicity and therapeutic failure. Platelet‐rich plasma (PRP) has been used for stimulating tissue regeneration, and mesenchymal stromal cells (MSCs) have demonstrated therapeutic properties in all phases of skin regeneration in cell therapy studies. Aims The objective of this study was to evaluate the therapeutic effects related to the use of a biomembrane composed of autologous MSCs and PRP on chronic wounds of diabetic patients (pre‐post pilot study). Patients/Methods Six diabetic patients with chronic wounds for more than 6 months were subjected to adipose tissue collection for isolation of MSCs, blood collection for PRP preparation, and topical administration of a biomembrane of MSCs and PRP on each chronic wound. The statistical difference regarding the evolution of ulcers was calculated by means of paired t test. Results There was granulation tissue formation starting from 7 days after topical application. Total re‐epithelialization occurred in 5 of the 9 lesions treated, and the mean wound healing rate (WHR) was 74.55% (±32.55%) after 90 days. No cicatricial hypertrophy or retraction was observed. Conclusion Mesenchymal stromal cells topical therapy associated with PRP is well‐tolerated and able to provide a reduction in ulcer area of diabetic chronic wounds.