Cells from adipose tissue fragments migrated and formed multicellular structures connecting different fragments in 3D cultures, suggesting chemotactic signaling between cells. RT-PCR showed the presence of mesenchymal stem cells and endothelial cells. A simple mechanical digestion procedure was used to prepare a stromal vascular fraction from canine adipose tissue for potential point-of-care applications in veterinary medicine.

1. RESULTS:
A high number of actively proliferating cells migrated out from
fragments and populated the plastic surface or the 3D matrix
In 3D cultures, outgrowing cells (OC) developed morphologically
complex multicellular structures connecting different fragments
(F). The formation of these structures suggests chemotactic
signaling between cells.
RT-PCR : MSCs and endothelial cells are present in cell cultures.
4X
20X 10X
2cm
OC
F
OC
F
F
OC
F
F
OC
Adipocytic differentiation
Osteocytic differentiation
ND*: not detected
Preparation of the stromal vascular fraction from adipose tissue
by mechanical digestion for point-of-care application in
veterinary medicine
GROLLI S., DI CASTRI A., CONTI V., MACCHI F., IVANOVSKA A., BASINI G., DEL BUE M., RAMONI R.
Dipartimento di Scienze Medico-Veterinarie, Via del Taglio 10, Università di Parma, 43126 Parma, Italy
OBJECTIVES: Adipose tissue derived Mesenchymal Stem Cells (AdMSCs) have a growing role in the therapy of a variety of animal’s
diseases, including wound healing, tendonitis, bone defects, osteoarticular pathologies. It is now accepted that fat tissue represents
indeed a source of mixed cell populations, as well as bioactive factors, able to contribute to the healing of injured tissues controlling
the inflammatory response, promoting the formation of blood vessels and modulating the different phases of tissue healing. The
term Stromal Vascular Fraction (SVF) has been introduced in recent years to indicate the mixed cell population (endothelial cells,
pericytes, smooth muscle cells, stromal progenitor cells, blood cells) isolated from fat tissue. The aim of this work was to evaluate
the feasibility of a simple procedure for the preparation of a SVF derived from subcutaneous and visceral fat in the dog, to be used
“point of care” ”, i.e. shortly after the collection of the biological sample and after a minimal manipulation.
MATERIALS AND METHODS:
A mechanical digestion was performed using a set of 6 mm
diameter sterile steel balls.
Cell cultures were maintained until passage four for cells
grown on plastic surface, or for 10-12 days for cells grown
inside the three-dimensional matrix.
Cell culture:
• PRP
• Thrombin
• DMEM
Methods:
1. on plastic surface
2. gel drops
3. 3D gel matrix
Conclusions: Although incomplete, the preliminary
characterization of the cells obtained by in-vitro culture of SVF,
let us hypothesize that the mixed cell population includes MSCs
maintaining their typical features. The safeguard of the tissue
architecture and the long-lasting maintenance of its vitality
guarantee a positive evaluation of the efficacy and efficiency of
the mechanical treatment.
≈ 2/3 hours
+PRP