scholarly journals Characterization and visualization of the liposecretion process taking place during ceiling culture of human mature adipocytes

2018 ◽  
Vol 234 (7) ◽  
pp. 10270-10280 ◽  
Author(s):  
Julie Anne Côté ◽  
Marie‐Frédérique Gauthier ◽  
Giada Ostinelli ◽  
Dannick Brochu ◽  
Kerstin Bellmann ◽  
...  
Keyword(s):  
Ceiling Culture

Low-dose radiation pretreatment improves survival of human ceiling culture-derived proliferative adipocytes (ccdPAs) under hypoxia via HIF-1 alpha and MMP-2 induction

2015 ◽  
Vol 463 (4) ◽  
pp. 1176-1183 ◽  
Author(s):  
Naoki Adachi ◽  
Yoshitaka Kubota ◽  
Kentarou Kosaka ◽  
Shinsuke Akita ◽  
Yoshitarou Sasahara ◽  
...  
Keyword(s):  
Low Dose ◽  
Low Dose Radiation ◽  
Ceiling Culture ◽  
Dose Radiation

scholarly journals Temporal Changes in Gene Expression Profile during Mature Adipocyte Dedifferentiation

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Julie Anne Côté ◽  
Frédéric Guénard ◽  
Julie Lessard ◽  
Marc Lapointe ◽  
Simon Biron ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Cellular Proliferation ◽  
Matrix Remodeling ◽  
Mature Adipocyte ◽  
Tissue Samples ◽  
Ceiling Culture ◽  
Isolated Adipocytes

Objective. To characterize changes in gene expression profile during human mature adipocyte dedifferentiation in ceiling culture.Methods. Subcutaneous (SC) and omental (OM) adipose tissue samples were obtained from 4 participants paired for age and BMI. Isolated adipocytes were dedifferentiated in ceiling culture. Gene expression analysis at days 0, 4, 7, and 12 of the cultures was performed using Affymetrix Human Gene 2.0 STvi arrays. Hierarchical clustering according to similarity of expression changes was used to identify overrepresented functions.Results. Four clusters gathered genes with similar expression between day 4 to day 7 but decreasing expression from day 7 to day 12. Most of these genes coded for proteins involved in adipocyte functions (LIPE,PLIN1,DGAT2,PNPLA2,ADIPOQ,CEBPA,LPL,FABP4,SCD,INSR, andLEP). Expression of several genes coding for proteins implicated in cellular proliferation and growth or cell cycle increased significantly from day 7 to day 12 (WNT5A,KITLG, andFGF5). Genes coding for extracellular matrix proteins were differentially expressed between days 0, 4, 7, and 12 (COL1A1,COL1A2, andCOL6A3,MMP1, andTGFB1).Conclusion. Dedifferentiation is associated with downregulation of transcripts encoding proteins involved in mature adipocyte functions and upregulation of genes involved in matrix remodeling, cellular development, and cell cycle.

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.

scholarly journals Fibrin glue increases the cell survival and the transduced gene product secretion of the ceiling culture-derived adipocytes transplanted in mice

2011 ◽  
Vol 43 (3) ◽  
pp. 161 ◽  
Author(s):  
Yasuyuki Aoyagi ◽  
Masayuki Kuroda ◽  
Sakiyo Asada ◽  
Hideaki Bujo ◽  
Shigeaki Tanaka ◽  
...  
Keyword(s):  
Cell Survival ◽  
Gene Product ◽  
Fibrin Glue ◽  
Ceiling Culture

Differentiation of 3T3-L1 preadipocytes is inhibited under a modified ceiling culture

2015 ◽  
Vol 39 (5) ◽  
pp. 638-645 ◽  
Author(s):  
Ziyi Song ◽  
Jia Cheng ◽  
Hao Yang ◽  
Yuefeng Li ◽  
Qian Gao ◽  
...  
Keyword(s):  
Ceiling Culture

Ceiling culture of mature human adipocytes: use in studies of adipocyte functions

2000 ◽  
Vol 164 (2) ◽  
pp. 119-128 ◽  
Author(s):  
HH Zhang ◽  
S Kumar ◽  
AH Barnett ◽  
MC Eggo
Keyword(s):  
Cell Biology ◽  
Morphological Changes ◽  
Culture Method ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Human Adipocytes ◽  
Necrosis Factor Alpha ◽  
Ceiling Culture ◽  
Confocal Immunofluorescence Microscopy ◽  
Cellular Expression ◽  
Adipocyte Cell

Adipocytes contain large lipid droplets in their cytoplasm. When cultured, they float on top of the medium, clump together, and do not gain equal and sufficient access to the medium. Morphological changes cannot be observed and the majority of adipocytes undergo cell lysis within 72 h of isolation. We have used a ceiling culture method for human mature adipocytes which uses their buoyant property to allow them to adhere to a floating glass surface, where they remain viable for several weeks. Using confocal immunofluorescence microscopy we showed the cellular expression and subcellular localization of leptin in ceiling-cultured adipocytes. The secretion of leptin was increased from ceiling cultures following tumour necrosis factor-alpha treatment. Proliferation of mature human adipocytes in serum-containing medium was demonstrated by incorporation of bromodeoxyuridine, 2% of adipocytes showing positive incorporation after 4 h labelling. Proliferation was also evident from the budding of daughter cells. Apoptosis in the ceiling cultures was increased by 48 h serum deprivation (30-35 vs 10-15% in the control) and was assayed by propidium iodide staining and terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick-end labelling. Lipolysis, analysed by liquid scintillation counting, was increased by forskolin (10 microM for 90 min) and lipogenesis, shown by autoradiography, was stimulated by insulin (10 and 100 nM for 4 h). These findings indicate that ceiling-cultured adipocytes maintain adipocyte-specific functions and that ceiling culture, which overcomes the shortcomings of adipocyte suspension culture, can be used to study adipocyte cell biology.

Ceiling culture-derived proliferative adipocytes retain high adipogenic potential suitable for use as a vehicle for gene transduction therapy

2011 ◽  
Vol 301 (1) ◽  
pp. C181-C185 ◽  
Author(s):  
Sakiyo Asada ◽  
Masayuki Kuroda ◽  
Yasuyuki Aoyagi ◽  
Yoshitaka Fukaya ◽  
Shigeaki Tanaka ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Adipogenic Differentiation ◽  
Marker Gene ◽  
Stromal Vascular Fraction ◽  
Marker Genes ◽  
Medicine Research ◽  
Ceiling Culture ◽  
The Difference ◽  
Adipogenic Marker

Adipose tissue is expected to provide a source of proliferative cells for regenerative medicine and cell-transplantation therapies using gene transfer manipulation. We have recently identified ceiling culture-derived proliferative adipocytes (ccdPAs) from the mature adipocyte fraction as cells suitable as a therapeutic gene vehicle because of their stable proliferative capacity. In this study, we examined the capability of adipogenic differentiation of the ccdPAs compared with stromal vascular fraction (SVF)-derived progenitor cells (adipose-derived stem cells, ASCs) with regard to their multipotential ability to be converted to another lineage and therefore their potential to be used for regenerative medicine research. After in vitro passaging, the surface antigen profile and the basal levels of adipogenic marker genes of the ccdPAs were not obviously different from those of the ASCs. However, the ccdPAs showed increased lipid-droplet accumulation accompanied with higher adipogenic marker gene expression after stimulation of differentiation compared with the ASCs. The higher adipogenic potential of the ccdPAs than the ASCs from the SVF was maintained for 42 days in culture. Furthermore, the difference in the adipogenic response was enhanced after partial stimulation without indomethacin. These results indicate that the ccdPAs retain a high adipogenic potential even after in vitro passaging, thus suggesting the commitment of ccdPAs to stable mature adipocytes after autotransplantation, indicating that they may have potential for use in regenerative and gene-manipulated medicine.

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