Isolation of mesenchymal stromal cells (MSCs) from adipose tissue is a relevant topic in regenerative medicine and cell therapy. MSCs have significant potential for treating various diseases due to their ability to differentiate, self-renew, and secrete biologically active molecules. However, the isolation process faces challenges like variations in tissue collection methods, quality, sample transportation, laboratory sterility, and incubation stability. Insufficient coverage of these issues in scientific publications complicates their resolution, particularly under the conditions in Ukraine.

The aim is to identify and discuss the key problems associated with the isolation of adipose-derived mesenchymal stromal cells, as well as the experimental search for potential solutions to overcome these obstacles for the optimization of the isolation methodology.

Materials and methods. A literature review was conducted using PubMed and Google Scholar, selecting key publications on isolation methods, MSCs characteristics, and good manufacturing practice (GMP) principles. Inclusion criteria: full-text articles on adipose MSCs isolation, comparison of enzymatic methods of isolation, GMP standardization, and cell characteristics. Following this, a pilot study was conducted using lipoaspirate and fragments of subcutaneous adipose tissue (SAT) with the patients’ consent. Processing in Zaporizhzhia State Medical and Pharmaceutical University’s GMP-compliant lab: mechanical grinding (for SAT), DPBS washing with antibiotics, enzymatic digestion (collagenase or trypsin) at 37 °C, 600 g centrifugation, filtration, DMEM / FBS cultivation. Viability was assessed in Goryaev chamber with trypan blue; passaging at 60–70 % confluency.

Results. Abdominal lipoaspirate yielded larger MSCs volumes with higher proliferation than thigh or excised SAT. Collagenase is considered the “gold standard” in terms of efficiency, but trypsin has been shown to be a cost-effective alternative with similar viability, adhesion, and differentiation (chondro-, osteo-, adipogenic). Cultivation with medium changes supported growth; passaging prevented senescence.

Conclusions. Optimization of the methodology for isolating mesenchymal stromal cells of adipogenic origin under GMP-compliant conditions can be achieved by using lipoaspirate from the abdominal area, using trypsinization as an effective and cost-effective alternative to collagenase for enzymatic isolation, and strictly adhering to cultivation, passage, and quality control protocols.