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Background Aims: Since initial methods were developed for isolating cells from adipose tissue, little has been done to improve mesenchymal stromal cell (MSC) yield. The aim of the present study was to isolate a population of MSC from lipoaspirate samples without tissue digestion and to assess the possibility of cryopreserving the freshly isolated cells.
Methods: A population of MSC was isolated from 13 patients' lipoaspirate samples by mechanical dissociation. Mechanically processed lipoaspirate adipose tissue (MPLA) cells were characterized after in vitro cell expansion by morphologic analysis, expression of MSC surface markers and differentiation assays.
Results: Mechanical dissociation yielded a large quantity of adherent MSC both after standard and vibro-assisted liposuction. Preservation of lipoaspirate samples at 4 degrees C for 1 or 2 days until the mechanical procedure did not change the MPLA cell content. It was possible to store freshly isolated MPLA cells by cryopreservation without loss of the MSC population. Adherent MPLA cells were negative for CD45 and CD31 and positive for CD34, CD105, CD44 and CD90. They also showed adipogenic, osteogenic and chondrogenic potentials similar to MSC populations from other sources as already described in the literature.
Conclusions: MSC can be isolated from human lipoaspirate samples by the mechanical procedure described in this study with a significant reduction in time and cost. Together with cryopreservation of freshly isolated MPLA cells, this has made it easier to harvest and store MSC for therapeutic applications such as soft-tissue augmentation and tissue engineering.
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http://dx.doi.org/10.3109/14653240902981144 | DOI Listing |
Stem Cell Res Ther
August 2025
Metabolic and Systemic Aspects of Oncogenesis for New Therapeutic Approaches (METSY), UMR 9018, CNRS, Gustave Roussy, Université Paris-Saclay, PR2, 114 Rue Edouard Vaillant, 94805, Villejuif, France.
Background: Under regular culture conditions, mesenchymal stem cells (MSCs) exhibit cytosolic calcium concentration oscillations (Ca oscillations), that change, especially in frequency, after the differentiation of the MSCs. Ca oscillations are known to encode important information in frequency and amplitude, ultimately controlling many cellular processes such as proliferation and differentiation. Previous studies evidenced that decreasing the frequency of Ca oscillations by physical means can facilitate osteodifferentiation of MSCs.
View Article and Find Full Text PDFMethods Mol Biol
May 2025
Institute for Cellular and Molecular Medicine, Department of Medical Immunology, and SAMRC Extramural Unit for Stem Cell Research and Therapy, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa.
Adipose-derived mesenchymal stromal/stem cells (AD-MSCs) offer a promising cell source for regenerative medicine due to their easy accessibility and potential application across a wide range of disorders. This chapter describes a standardized enzymatic isolation protocol to extract AD-MSCs from both solid adipose tissue and lipoaspirate. Adipose tissue samples are initially washed with phosphate-buffered saline (PBS) to remove blood contaminants.
View Article and Find Full Text PDFCells
April 2025
Department of Plastic, Hand and Reconstructive Surgery, University Hospital Regensburg, 93053 Regensburg, Germany.
Manual processing of lipoaspirate can enhance stem cell concentration, thereby improving the take rate, which still represents a major challenge in autologous fat transfer. However, since the preparation consists of many manual steps that are difficult to standardize, we investigated the influence of residual tumescent solution on the macroscopic and microscopic outcome of the mechanically processed lipoaspirate. Additionally, we investigated whether sedimentation followed by vacuum filtration of the aqueous phase could accelerate processing by replacing the initial centrifugation step.
View Article and Find Full Text PDFBackground: Mechanical processing of lipoaspirate (LA) produces a stromal vascular fraction (SVF) without enzymatic digestion for use in aesthetic, surgical, and regenerative applications. We recently presented novel device technologies that increased mesenchymal stem cell (MSC) content relative to standard nanofat (NF) processing.
Objectives: Here, we introduce a third technology designed to enhance fluid shear forces and explore the impact of mechanical processing on regenerative potential in vitro.
Front Cell Dev Biol
February 2025
Department of Orthopaedic Surgery, Experimental Orthopaedics, Centre for Medical Biotechnology (ZMB/Biopark 1), University of Regensburg, Regensburg, Germany.
Introduction: Orthobiologics, such as autologous nanofat, are emerging as a potential treatment option for osteoarthritis (OA), a common degenerative joint causing pain and disability in the elderly. Nanofat, a minimally processed human fat graft rich in stromal vascular fraction (SVF) secretory factors, has shown promise in relieving pain. This study aimed to elucidate the molecular mechanisms underlying nanofat treatment of OA-affected cells and compare two filtration systems used for nanofat preparation.
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