The Effect of Centrifugation Condition on Mature Adipocytes and Adipose Stem Cell Viability

Abstract

Different researchers have recommended different lipoaspirate centrifugation speeds and times, probably due to the limits in fat cell viability assays. We assessed fat cell viability using a fluorescein diacetate and propidium iodide (FDA-PI) stain and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay after harvesting syringe liposuction and spun with different centrifugation speeds to determine the optimal conditions. Lipoaspirates, harvested from 13 donors, were transferred into a centrifuge tube and spun at 1000, 3000, and 4000 rpm for 3 minutes. Mature adipocytes and adipose stem cells were isolated and tested with a direct counting of FDA-PI-stained cells under fluorescence microscope and XTT assay. We incubated adipocytes and adipose stem cells for 1 day and 3 days, and we compared both of them with fresh samples to evaluate the influence of culturing condition on fat cell viability. Centrifugation speeds from 1000 rpm to 4000 rpm for 3 minutes showed no change in the percentage of adipocytes and adipose stem cell viability not only in the fresh samples but also in the cultured samples (1 day and 3 days). Centrifugation speeds under 4000 rpm do not change the percentage of fat cell viability. To differentiate viable cells from dying or dead mature adipocytes and oil accurately, combinations of viability tests are essential.

Lull pgm system: A suitable technique to improve the regenerative potential of autologous fat grafting

“…Total DNA extraction of the oily adipocyte upper fraction was performed using the samples isolated from six patients (triplicate measurements, n = 6). The amount of DNA is representative of the number of mature adipocytes of each sample because only this type of cells are present in the upper layer . These values were used to compare the relative amount of the adipocytes contained in the processed lipoaspirates.…”

“…The amount of DNA is representative of the number of mature adipocytes of each sample because only this type of cells are present in the upper layer. 25 These values were used to compare the relative amount of the adipocytes contained in the processed lipoaspirates. No significant difference among the Coleman (C), decantation (D), and Lull (L) groups was observed, although it can be noticed that the highest mean value was obtained with the Lull group (Figure 2A).…”

The Selective Centrifugation Ensures a Better In Vitro Isolation of ASCs and Restores a Soft Tissue Regeneration In Vivo

“…To the best of our knowledge, there are few publications of a prospective and controlled study investigating the use of lipofilling breast volumetry in the field of breast reconstruction. Careful analysis of the literature highlights the presence of several approaches to fat graft differing in harvesting, centrifugation or infiltration methods [32,33,34,35,36]. In particular, some authors strongly support the potential of stem cells for the purpose of increasing fat graft survival, indicating the enrichment of lipofilling with stem cells as the best alternative to major tissue augmentation [37,38].…”

Comparison of Methods in Studies of Cell Death Mechanisms

“…On the other hand, Son et al proved that centrifugation with frequencies in the range of 1000-4000 rpm applied for 3 min has no influence on percentage of adipocytes and adipose stem cell viability [10]. Therefore, we postulate that centrifugation itself or further resuspension of cell's pellet may lead to cell damage and increase percentage of the necrotic cells in population.…”

“…Cell culture parameters, i.e. chosen variant of cell harvesting, either by mechanical scrapping or by enzymes and centrifugation may lead to false positive or false negative results [4][5][6][7][8][9][10].…”