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Fett bietet neben seinem volumisierenden Effekt ein reichhaltiges Reservoir fur regenerative Zellen, die eine wichtige Rolle fur den Einbau und das Uberleben eines Fettgrafts spielen. Wir haben den Unterschied zwischen dermalem weisem Fettgewebe (dWAT) und subkutanem weisem Fettgewebe (sWAT) als auch die Auswirkungen der maschinellen und manuellen Fettabnahme mit verschiedenen Kanulen- bzw. Perforationsgrosen auf die Viabilitat der Adipozyten und der aus dem Fettgewebe isolierten stromalen vaskularen Fraktion (SVF) untersucht. Zusatzlich verglichen wir, ob eine hohe Zentrifugation sowie die Tumeszenzlosung einen Effekt auf diese Zellen haben. Es zeigte sich, dass mit einer dWAT-Kanule mehr tote Adipozyten hervorgebracht werden, jedoch mit einer hoheren SVF-Zellausbeute. Die aus dWAT isolierten SVF-Zellen zeigten eine leicht erhohte Zelldichte und metabolische Aktivitat nach Isolierung im Vergleich zu aus sWAT isolierten SVF-Zellen. Im Gegensatz zur 2‑mm-Kanule war die 1‑mm-Kanule schonender in der Fettentnahme und brachte signifikant viable SVF-Zellen hervor. Einen negativen Effekt auf das Uberleben der Adipozyten zeigte unser Versuch, Fettgrafts mittels 2 Spritzen und Sieb herzustellen. Das Waschen mit NaCl, um die Tumeszenzlosung zu entfernen, resultierte in weniger toten Zellen im Liposuktionsmaterial. Auch die vielerorts verwendete Zentrifugation des Liposuktionsmaterials hatte einen negativen Einfluss auf das Uberleben der Adipozyten. Ein Eigenfetttransfer besteht aus vielen einzelnen Schritten. Das Ergebnis hangt nicht nur von der Wahl der Kanulen, sondern auch von der Verarbeitung und Reinjektionstechnik des gewonnenen Gewebes ab. Ziel ist, jeden einzelnen Verarbeitungsschritt zu verbessern, um dem transplantierten Gewebe die bestmoglichen Uberlebenschancen zu bieten und dadurch das Endergebnis zu optimieren.

The prerequisite for a successful clinical use of autologous adipose-tissue-derived cells is the highest possible regenerative potential of the applied cell population, the stromal vascular fraction (SVF). Current isolation methods depend on high enzyme concentration, lysis buffer, long incubation steps and mechanical stress, resulting in single cell dissociation. The aim of the study was to limit cell manipulation and obtain a derivative comprising therapeutic cells (microtissue-SVF) without dissociation from their natural extracellular matrix, by employing a gentle good manufacturing practice (GMP)-grade isolation. The microtissue-SVF yielded larger numbers of viable cells as compared to the improved standard-SVF, both with low enzyme concentration and minimal dead cell content. It comprised stromal tissue compounds (collagen, glycosaminoglycans, fibroblasts), capillaries and vessel structures (CD31+, smooth muscle actin+). A broad range of cell types was identified by surface-marker characterisation, including mesenchymal, haematopoietic, pericytic, blood and lymphatic vascular and epithelial cells. Subpopulations such as supra-adventitial adipose-derived stromal/stem cells and endothelial progenitor cells were significantly more abundant in the microtissue-SVF, corroborated by significantly higher potency for angiogenic tube-like structure formation in vitro. The microtissue-SVF showed the characteristic phenotype and tri-lineage mesenchymal differentiation potential in vitro and an immunomodulatory and pro-angiogenic secretome. In vivo implantation of the microtissue-SVF combined with fat demonstrated successful graft integration in nude mice. The present study demonstrated a fast and gentle isolation by minor manipulation of liposuction material, achieving a therapeutically relevant cell population with high vascularisation potential and immunomodulatory properties still embedded in a fraction of its original matrix.

Positive Effekte der extrakorporalen Stoswellentherapie (ESWT) konnten zuvor auf Wundheilung, Nervenregeneration und Gefasneubildung bewiesen werden. In dieser Studie haben wir die radiale ESWT zur Gesichtsrejuvenation untersucht, wobei bereits die direkte ESWT-Behandlung erste Wirkung gezeigt hat. Fur eine gesteigerte Volumisierung wurde zusatzlich Eigenfett mit ESWT-Vorbehandlung des Spender-und Empfangerareals appliziert. Die Vorbehandlung des Empfangerareals mit ESWT ist fur das Anwachsen des Transplantats notwendig, da mittels ESWT die Angiogenese und somit ein erfolgreicher Einbau der Grafts im Zielgewebe erfolgen kann.

A highly interesting source for adult stem cells is adipose tissue, from which the stromal vascular fraction (SVF)-a heterogeneous cell population including the adipose-derived stromal/stem cells-can be obtained. To enhance the regenerative potential of freshly isolated SVF cells, low-level light therapy (LLLT) was used. The effects of pulsed blue (475 nm), green (516 nm), and red (635 nm) light from light-emitting diodes applied on freshly isolated SVF were analysed regarding cell phenotype, cell number, viability, adenosine triphosphate content, cytotoxicity, and proliferation but also osteogenic, adipogenic, and proangiogenic differentiation potential. The colony-forming unit fibroblast assay revealed a significantly increased colony size after LLLT with red light compared with untreated cells, whereas the frequency of colony-forming cells was not affected. LLLT with green and red light resulted in a stronger capacity to form vascular tubes by SVF when cultured within 3D fibrin matrices compared with untreated cells, which was corroborated by increased number and length of the single tubes and a significantly higher concentration of vascular endothelial growth factor. Our study showed beneficial effects after LLLT on the vascularization potential and proliferation capacity of SVF cells. Therefore, LLLT using pulsed light-emitting diode light might represent a new approach for activation of freshly isolated SVF cells for direct clinical application.

One of the mainstays of facial rejuvenation strategies is volume restoration, which can be achieved by autologous fat grafting. In our novel approach, we treated the adipose tissue harvest site with extracorporeal shock wave therapy (ESWT) in order to improve the quality of the regenerative cells in situ. The latter was demonstrated by characterizing the cells of the stromal vascular fraction (SVF) in the harvested liposuction material regarding cell yield, adenosine triphosphate (ATP) content, proliferative capacity, surface marker profile, differentiation potential and secretory protein profile. Although the SVF cell yield was only slightly enhanced, viability and ATP concentration of freshly isolated cells as well as proliferation doublings after 3 weeks in culture were significantly increased in the ESWT compared with the untreated group. Likewise, cells expressing mesenchymal and endothelial/pericytic markers were significantly elevated concomitant with an improved differentiation capacity towards the adipogenic lineage and enhancement in specific angiogenic proteins. Hence, in situ ESWT might be applied in the future to promote cell fitness, adipogenesis and angiogenesis within the fat graft for successful facial rejuvenation strategies with potential long-term graft survival.

Background aims Lipedema is a hormone-related disease of women characterized by enlargement of the extremities caused by subcutaneous deposition of adipose tissue. In healthy patients application of autologous adipose tissue–derived cells has shown great potential in several clinical studies for engrafting of soft tissue reconstruction in recent decades. The majority of these studies have used the stromal vascular fraction (SVF), a heterogeneous cell population containing adipose-derived stromal/stem cells (ASC), among others. Because cell identity and regenerative properties might be affected by the health condition of patients, we characterized the SVF cells of 30 lipedema patients in comparison to 22 healthy patients. Methods SVF cells were analyzed regarding cell yield, viability, adenosine triphosphate content, colony forming units and proliferative capacity, as well as surface marker profile and differentiation potential in vitro. Results Our results demonstrated a significantly enhanced SVF cell yield isolated from lipedema compared with healthy patients. In contrast, the adipogenic differentiation potential of SVF cells isolated from lipedema patients was significantly reduced compared with healthy patients. Interestingly, expression of the mesenchymal marker CD90 and the endothelial/pericytic marker CD146 was significantly enhanced when isolated from lipedema patients. Discussion The enhanced number of CD90+ and CD146+ cells could explain the increased cell yield because the other tested surface marker were not reduced in lipedema patients. Because the cellular mechanism and composition in lipedema is largely unknown, our findings might contribute to a better understanding of its etiology.

Background Cell-based therapies with autologous adipose tissue–derived cells have shown great potential in several clinical studies in the last decades. The majority of these studies have been using the stromal vascular fraction (SVF), a heterogeneous mixture of fibroblasts, lymphocytes, monocytes/macrophages, endothelial cells, endothelial progenitor cells, pericytes and adipose-derived stromal/stem cells (ASC) among others. Although possible clinical applications of autologous adipose tissue–derived cells are manifold, they are limited by insufficient uniformity in cell identity and regenerative potency. Methods In our experimental set-up, low-energy extracorporeal shock wave therapy (ESWT) was performed on freshly obtained human adipose tissue and isolated adipose tissue SVF cells aiming to equalize and enhance stem cell properties and functionality. Results After ESWT on adipose tissue we could achieve higher cellular adenosine triphosphate (ATP) levels compared with ESWT on the isolated SVF as well as the control. ESWT on adipose tissue resulted in a significantly higher expression of single mesenchymal and vascular marker compared with untreated control. Analysis of SVF protein secretome revealed a significant enhancement in insulin-like growth factor (IGF)-1 and placental growth factor (PLGF) after ESWT on adipose tissue. Discussion Summarizing we could show that ESWT on adipose tissue enhanced the cellular ATP content and modified the expression of single mesenchymal and vascular marker, and thus potentially provides a more regenerative cell population. Because the effectiveness of autologous cell therapy is dependent on the therapeutic potency of the patient's cells, this technology might raise the number of patients eligible for autologous cell transplantation.