scholarly journals Evaluation of Three Devices for the Isolation of the Stromal Vascular Fraction from Adipose Tissue and for ASC Culture: A Comparative Study

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Jonathan Rodriguez ◽  
Anne-Sophie Pratta ◽  
Nacira Abbassi ◽  
Hugo Fabre ◽  
Fanny Rodriguez ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Adipose Tissue ◽  
Medical Devices ◽  
Stromal Cells ◽  
Genetic Stability ◽  
Stromal Vascular Fraction ◽  
Reference Method ◽  
Proliferative Potential ◽  
Population Doublings ◽  
Multilineage Potential

Adipose-derived stem/stromal cells (ASCs) reside in the stromal vascular fraction (SVF) of adipose tissue (AT) and can be easily isolated. However, extraction of the SVF from lipoaspirate is a critical step in generating ASC, and semiautomated devices have been developed to enhance the efficacy and reproducibility of the outcomes and to decrease manipulation and contamination. In this study, we compared the reference method used in our lab for SVF isolation from lipoaspirate, with three medical devices: GID SVF-1™, Puregraft™, and Stem.pras®. Cell yield and their viability were evaluated as well as their phenotype with flow cytometry. Further on, we determined their proliferative potential using population doublings (PD), PD time (PDT), and clonogenicity assay (CFU-F). Finally, we checked their genetic stability using RT-qPCR for TERT mRNA assay and karyotyping as well as their multilineage potential including adipogenic, chondrogenic, and osteogenic differentiation. Our results demonstrate that all the devices allow the production of SVF cells with consistent yield and viability, in less time than the reference method. Expanded cells from the four methods showed no significant differences in terms of phenotype, proliferation capabilities, differentiation abilities, and genetic stability.

Adipose-Derived Stem/Stromal Cells, Stromal Vascular Fraction, and Microfragmented Adipose Tissue

2022 ◽  
pp. 47-61
Author(s):  
Enrico Ragni ◽  
Marco Viganò ◽  
Paola De Luca ◽  
Edoardo Pedrini ◽  
Laura de Girolamo
Keyword(s):  
Adipose Tissue ◽  
Stromal Cells ◽  
Stromal Vascular Fraction

scholarly journals A Novel Method to Optimize Autologous Adipose Tissue Recovery with Extracellular Matrix Preservation

Processes ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 88
Author(s):  
Ilaria Roato ◽  
Federico Mussano ◽  
Simone Reano ◽  
Filippo Boriani ◽  
Andrea Margara ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Extracellular Matrix ◽  
Adipose Tissue ◽  
Regenerative Medicine ◽  
Stromal Vascular Fraction ◽  
Fold Increase ◽  
Human Adipose Tissue ◽  
Promising Tool ◽  
Filter Size ◽  
Tissue Recovery

This work aims to characterize a new method to recover low-manipulated human adipose tissue, enriched with adipose tissue-derived mesenchymal stem cells (ATD-MSCs) for autologous use in regenerative medicine applications. Lipoaspirated fat collected from patients was processed through Lipocell, a Class II-a medical device for dialysis of adipose tissue, by varying filter sizes and washing solutions. ATD-MSC yield was measured with flow cytometry after stromal vascular fraction (SVF) isolation in fresh and cultured samples. Purification from oil and blood was measured after centrifugation with spectrophotometer analysis. Extracellular matrix preservation was assessed through hematoxylin and eosin (H&E) staining and biochemical assay for total collagen, type-2 collagen, and glycosaminoglycans (GAGs) quantification. Flow cytometry showed a two-fold increase of ATD-MSC yield in treated samples in comparison with untreated lipoaspirate; no differences where reported when varying filter size. The association of dialysis and washing thoroughly removed blood and oil from samples. Tissue architecture and extracellular matrix integrity were unaltered after Lipocell processing. Dialysis procedure associated with Ringer’s lactate preserves the proliferation ability of ATD-MSCs in cell culture. The characterization of the product showed that Lipocell is an efficient method for purifying the tissue from undesired byproducts and preserving ATD-MSC vitality and extracellular matrix (ECM) integrity, resulting in a promising tool for regenerative medicine applications.

The Development of Facial Lipofilling from a Historical Point of View

2019 ◽  
Vol 35 (04) ◽  
pp. 358-367 ◽  
Author(s):  
Joris A. van Dongen ◽  
Joeri van Boxtel ◽  
Martin C. Harmsen ◽  
Hieronymus P. Stevens
Keyword(s):  
Adipose Tissue ◽  
Stromal Cells ◽  
Stromal Vascular Fraction ◽  
Point Of View ◽  
Congenital Defects ◽  
Skin Rejuvenation ◽  
Adipose Derived Stromal Cells ◽  
Enzymatic Isolation ◽  
The Face ◽  
Mechanical Isolation

AbstractLipofilling, the transplantation of adipose tissue, has already been used since the end of the 19th century. For decades, lipofilling was used to restore loss of volume due to aging, trauma, or congenital defects. Later on, the indications for the use of lipofilling expanded by treating aged skin, scars, and improving wound healing. The expansion was caused by the discovery of adipose derived stromal cells (ASCs) in adipose tissue and the development of very fine harvesting and injection cannulas which made it possible to inject small adipose tissue particles in small volume areas, such as the face. ASCs are multipotent stromal cells which reside in the stromal vascular fraction (SVF) of adipose tissue and are able to differentiate in multiple cell lineages and secrete a plurality of growth factors with regenerative potentials. The discovery of ASCs led toward more experimental cell-based therapies, that is, ASCs or SVF isolated by means of enzymatic isolation procedures. Later on, enzymatic isolation procedures were forbidden in many countries by legislation and were replaced by mechanical isolation procedures, such as the Nanofat and Fractionation of Adipose Tissue (FAT) procedures. The Nanofat procedure has been extensively investigated, especially as treatment for skin rejuvenation in the face. Though, substantial evidence is lacking for using facial lipofilling or any therapeutic component, that is, ASCs or SVF for skin rejuvenation to date. In contrast, facial lipofilling to restore loss of volume seems to be promising.

scholarly journals Infusing Mesenchymal Stromal Cells into Porcine Kidneys during Normothermic Machine Perfusion: Intact MSCs Can Be Traced and Localised to Glomeruli

2019 ◽  
Vol 20 (14) ◽  
pp. 3607 ◽  
Author(s):  
Merel Pool ◽  
Tim Eertman ◽  
Jesus Sierra Parraga ◽  
Nils ’t Hart ◽  
Marieke Roemeling-van Rhijn ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Adipose Tissue ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Human Adipose Tissue ◽  
Machine Perfusion ◽  
Normothermic Machine Perfusion ◽  
Glomerular Capillaries

Normothermic machine perfusion (NMP) of kidneys offers the opportunity to perform active interventions, such as the addition of mesenchymal stromal cells (MSCs), to an isolated organ prior to transplantation. The purpose of this study was to determine whether administering MSCs to kidneys during NMP is feasible, what the effect of NMP is on MSCs and whether intact MSCs are retained in the kidney and to which structures they home. Viable porcine kidneys were obtained from a slaughterhouse. Kidneys were machine perfused during 7 h at 37 °C. After 1 h of perfusion either 0, 105, 106 or 107 human adipose tissue derived MSCs were added. Additional ex vivo perfusions were conducted with fluorescent pre-labelled bone-marrow derived MSCs to assess localisation and survival of MSCs during NMP. After NMP, intact MSCs were detected by immunohistochemistry in the lumen of glomerular capillaries, but only in the 107 MSC group. The experiments with fluorescent pre-labelled MSCs showed that only a minority of glomeruli were positive for infused MSCs and most of these glomeruli contained multiple MSCs. Flow cytometry showed that the number of infused MSCs in the perfusion circuit steeply declined during NMP to approximately 10%. In conclusion, the number of circulating MSCs in the perfusate decreases rapidly in time and after NMP only a small portion of the MSCs are intact and these appear to be clustered in a minority of glomeruli.

Human mesenchymal adipose stromal cells from mature adipocyte fraction

2010 ◽  
Vol 5 (1) ◽  
pp. 47-58 ◽  
Author(s):  
Aušra Unguryte ◽  
Eiva Bernotiene ◽  
Algirdas Venalis
Keyword(s):  
Adipose Tissue ◽  
Stromal Cells ◽  
Stromal Vascular Fraction ◽  
Culture Method ◽  
Control Culture ◽  
Mature Adipocyte ◽  
Ceiling Culture ◽  
Bottom Cell ◽  
Cell Fractions ◽  
Proliferation Potential

AbstractMore effective techniques should be employed for isolation of human mesenchymal stromal cells derived from adipose tissue (ADSC), seeking to make adipose tissue biopsies smaller in volume and thus less invasive. In this study, we compared properties of ADSC isolated by several different methods from the same samples of adipose tissue in order to enhance yields of potential ADSC. The mature adipocyte fraction was investigated using the ceiling culture method, including both ceiling and bottom cell fractions, and the control culture method with standard amount of medium. The results were also compared using the stromal vascular fraction from the same samples. The most efficient was the bottom cell population isolated from the mature adipocyte fraction by ceiling culture method. These cells readily differentiated into osteogenic, adipogenic and chondrogenic lineages and, similar to stromal vascular fraction cells, displayed high proliferation potential. Cultures of mature adipocyte fractions with standard amount of medium were considerably less effective. Mature adipocyte fractions yields large quantities of adipose-derived stem cells that have properties comparable with stromal vascular fraction cells suitable for tissue regeneration, especially when only small biopsies can be taken.

236. Characterising the stem cell activity of human endometrial epithelial and stromal cells

2005 ◽  
Vol 17 (9) ◽  
pp. 93 ◽  
Author(s):  
C. E. Gargett ◽  
R. M. Zillwood ◽  
K. E. Schwab ◽  
S. Z. Naqvi
Keyword(s):  
Stem Cell ◽  
Stromal Cells ◽  
Cell Activity ◽  
Human Endometrium ◽  
Proliferative Potential ◽  
Epithelial Stem Cells ◽  
Regenerative Capacity ◽  
Self Renewal ◽  
Endometrial Cells ◽  
Population Doublings

The human endometrium has remarkable regenerative capacity. It contains rare clonogenic epithelial and stromal cells.1 The aims of this study were to compare the stem cell properties, self renewal, differentiation and proliferative potential, of endometrial cells initiating large clones (HPP-CFU, high proliferative potential–colony forming units) with those initiating small clones (LPP-CFU, low PP-CFU). Endometrial tissue obtained from 12 ovulating women undergoing hysterectomy was dissociated into single cell suspensions and epithelial and stromal cells were cultured at clonal density (8–20cells/cm2). Individual clones were harvested and serially recloned to measure self-renewal, serially passaged to assess proliferative potential and cultured in various media to assess differentiation. Secondary stromal clones were characterized for mesenchymal stem cell (MSC) markers by flow cytometry. Epithelial HPP-CFU serially cloned significantly more times than epithelial LPP-CFU (P<0.005, Table1) indicating greater self renewal capacity. Similarly, stromal HPP-CFU demonstrated greater self renewal activity compared with stromal LPP-CFU (P<0.005, Table1). Serially passaged epithelial HPP-CFU underwent significantly more population doublings (PD) before senescence than epithelial LPP-CFU (P<0.05, Table1). Likewise, stromal HPP-CFU underwent a higher number of PD compared with stromal LPP-CFU (P<0.005, Table1). Stromal HPP-CFU expressed CD29, CD44, CD73, CD90, CD105 and CD146, and were negative for haemopoietic and endothelial markers, CD45, CD34 and CD31. Stromal HPP-CFU differentiated into adipocytes, smooth muscle cells, osteoblasts and chondrocytes when cultured in appropriate differentiation media. Our studies of human endometrial cells demonstrate that the rare epithelial and stromal HPP-CFU exhibit self-renewal and high proliferative potential. Furthermore stromal HPP-CFU express markers typical of bone marrow MSC and differentiate into four mesenchymal lineages. These data suggest that epithelial and stromal HPP-CFU, but not LPP-CFU, have characteristic properties of adult epithelial stem cells and MSC respectively and are likely responsible for the remarkable cyclical, regenerative capacity of human endometrium.

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