Human adipose stromal vascular cell delivery in a fibrin spray

Abstract

Background Adipose tissue represents a practical source of autologous mesenchymal stromal cells (MSC) and vascular-endothelial progenitor cells, available for regenerative therapy without in vitro expansion. One of the problems confronting the therapeutic application of such cells is how to immobilize them at the wound site. Here, we evaluated in vitro the growth and differentiation of human adipose stromal vascular fraction (SVF) cells after delivery using a fibrin spray system. Methods SVF cells were harvested from four human adult patients undergoing elective abdominoplasty using the LipiVage™ system. After collagenase digestion, mesenchymal and endothelial progenitor cells (pericytes, supra-adventitial stromal cells, endothelial progenitors) were quantified by flow cytometry before culture. SVF cells were applied to culture vessels using the Tisseel™ fibrin spray system. SVF cell growth and differentiation was documented by immunofluorescence staining and photomicrography. Results SVF cells remained viable following application and were expanded up to three weeks, when they reached confluence and adipogenic differentiation. Under angiogenic conditions, SVF cells formed endothelial (vWF+, CD31+ and CD34+) tubules surrounded by CD146+ and α-SMA+ perivascular/stromal cells. Discussion Human adipose tissue is a rich source of autologous stem cells, which are readily available for regenerative applications such as wound healing, without in vitro expansion. Our results indicate that mesenchymal and endothelial progenitor cells, prepared in a closed system from unpassaged lipoaspirate samples, retain their growth and differentiation capacity when applied and immobilized on a substrate using a clinically approved fibrin sealant spray system.

Mesenchymal stem cell secretome and regenerative therapy after cancer

“…Several studies have investigated the effects of BM-MSC and HSC co-transplantation to facilitate engraftment or reduce graft-versus-host disease into patients treated for hematopoietic malignancies [16, 22, 23]. Autologous BM-MSC were also delivered in a fibrin spray to accelerate wound healing in patients with acute wounds including skin cancer surgery-induced lesions [24], and our group has recently validated in vitro an analogous strategy using unpassaged adipose-derived MSC [25]. Intrabone and systemic delivery of MSC has been tested in a multiple myeloma animal model for simultaneous inhibition of tumor growth and regeneration of bone lesions [26].…”

Fibroblast-Negative CD34-Negative Cells from Human Adipose Tissue Contain Mesodermal Precursors for Endothelial and Mesenchymal Cells

“…4B); additionally, CD45 -FIB -CD34 -and CD45 -FIB -CD34 + cells expressed Tie2, KDR and Flt1 at similar levels (approximately 1.0%, 0.5% and 5%, respectively), but CD9 and CD90 had higher expression levels in the CD34 + cell population (approximately 85% and 15%, respectively) than in its counterpart CD34 -cell subset (approximately 30% and 2%, respectively). To assess the ability of these primitive CD34 -cells to generate endothelial cells in vitro, freshly isolated CD45 -FIB - 16 16 CD34 -cells were plated on uncoated 6-well plastic plates, and after 7 days, the nonadherent cells (Fig. 4C) were re-plated in replicate fibronectin/collagen-coated plates in EGM; in some experiments, the non-adherent cells were grown in semisolid medium.…”

Adipose Tissue-Derived Mesenchymal Stem Cells: A Novel Therapy in Orthopedics

“…SVF has been used as a source of cells useful for tissue regeneration. The most common isolation technique to obtain SVF is the enzymatic digestion which consists of washing, treatment with enzymes, erythrocyte lysis, and cryopreservation or culture expansion [55][56][57][58][59][60][61][62][63][64]. This technique is costly and might jeopardize the safety as well as the efficacy of the lipoaspirates [4,65].…”