A New Organotypic Culture of Adipose Tissue Fragments Maintains Viable Mature Adipocytes for a Long Term, Together with Development of Immature Adipocytes and Mesenchymal Stem Cell-Like Cells

Sonoda, Emiko; Aoki, Shigehisa; Uchihashi, Kazuyoshi; Soejima, Hidenobu; Kanaji, Sachiko; Izuhara, Kenji; Satoh, Seiji; Fujitani, Noboru; Sugihara, Hajime; Toda, Shuji

doi:10.1210/en.2008-0525

Cited by 37 publications

(67 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…4), both preadipocytes and MSCs actively regenerate at the peripheral zone of the fragments, whereas they are not produced at the central zone. 9 In fact, cellular growth at the peripheral zone is extensively higher than that at the central zone (Fig. 7A).…”

Section: Organotypic Culture As a Model For Tissue Regeneration And Omentioning

confidence: 90%

“…On the basis of this fact, organotypic culture of tissue fragments seems to be a promising model for investigating tissue regeneration and remodeling in vitro. However, only several organotypic cultures of tissue fragments, including thyroid, 27,28 brain 31,32 and adipose tissues, 9 are successfully established. Thus, various tissues other than these tissues above should be applied to organotypic culture system.…”

Section: Organotypic Culture As a Model For Tissue Regeneration And Omentioning

confidence: 99%

“…9 Our method named "adipose tissue-organotypic culture" maintains the unilocular structure and active function of mature adipocytes for more than 4 weeks. Furthermore, both preadipocytes and MSCs develop actively around adipose tissue fragments.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Adipose tissue-organotypic culture system as a promising model for studying adipose tissue biology and regeneration

et al. 2009

Self Cite

View full text Add to dashboard Cite

Adipose tissue consists of mature adipocytes, preadipocytes and mesenchymal stem cells (MSCs), but a culture system for analyzing their cell types within the tissue has not been established. We have recently developed "adipose tissue-organotypic culture system" that maintains unilocular structure, proliferative ability and functions of mature adipocytes for a long term, using three-dimensional collagen gel culture of the tissue fragments. In this system, both preadipocytes and MSCs regenerate actively at the peripheral zone of the fragments. Our method will open up a new way for studying both multiple cell types within adipose tissue and the cell-based mechanisms of obesity and metabolic syndrome. Thus, it seems to be a promising model for investigating adipose tissue biology and regeneration. In this article, we introduce adipose tissue-organotypic culture, and propose two theories regarding the mechanism of tissue regeneration that occurs specifically at peripheral zone of tissue fragments in vitro.

show abstract

Section: Organotypic Culture As a Model For Tissue Regeneration And Omentioning

confidence: 90%

Section: Organotypic Culture As a Model For Tissue Regeneration And Omentioning

confidence: 99%

See 1 more Smart Citation

Adipose tissue-organotypic culture system as a promising model for studying adipose tissue biology and regeneration

et al. 2009

Self Cite

View full text Add to dashboard Cite

show abstract

“…Mouse Primary Adipocyte Culture-Primary culture of epididymal adipose tissue was performed as described previously (17,18). Briefly, ϳ0.2 g of the epididymal adipose tissue from each mouse was rinsed with DMEM:F-12, minced to achieve pieces of ϳ0.5 mm in diameter, suspended in collagen matrix, and plated in 1-ml volumes in a six-well plate (18).…”

Section: Methodsmentioning

confidence: 99%

Cyclooxygenase-2 Deficiency Attenuates Adipose Tissue Differentiation and Inflammation in Mice

Ghoshal

Trivedi

Graf

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Obesity is associated with a variety of disorders and is a significant health problem in developed countries. One factor controlling the level of adiposity is the differentiation of cells into adipocytes. Adipocyte differentiation requires expression of peroxisome proliferator-activated receptor ␥ (PPAR␥), which is activated by ligands to regulate expression of genes involved in adipocyte differentiation. Although 15-deoxy-⌬(12,14)-prostaglandin (PG) J 2 (15d-PGJ(2)) has long been known to be a potent activator of PPAR␥, the importance of its synthesis in adipose tissue in vivo is not clear. The current study utilized mice deficient in cyclooxygenase-2 (COX-2) to examine the role of COX-2-derived PGs as in vivo modulators of adiposity. As compared with strain-and age-matched wildtype controls, the genetic deficiency of COX-2 resulted in a significant reduction in total body weight and percent body fat. Although there were no significant differences in food consumption between groups, COX-2-deficient mice showed increased metabolic activity. Epididymal adipose tissue from wild-type mice produced a significantly greater level of 15d-PGJ(2), as compared with adipose tissue isolated from mice deficient in COX-2. Furthermore, production of the precursor required for 15d-PGJ(2) formation, PGD(2), was also significantly reduced in COX-2-deficient adipose tissue. The expression of markers for differentiated adipocytes was significantly reduced in adipose tissue from COX-2-deficient mice, whereas preadipocyte marker expression was increased. Macrophagedependent inflammation was also significantly reduced in adipose tissue of COX-2-deficient mice. These findings suggest that reduced adiposity in COX-2-deficient mice results from attenuated PPAR␥ ligand production and adipocyte differentiation.In developed countries, obesity is a significant health problem that is becoming more prevalent. The increase in adiposity, or the percentage of total body mass comprised of lipid stores, correlates with the development of a variety of disorders including, glucose intolerance, dyslipidemia, and hypertension (1). Recent evidence suggests that in addition to being an energy storage depot, adipose tissue may contribute to disease by producing adipokines, chemokines, and other bioactive substances (2). The level of adiposity is regulated by the amount of lipid taken into individual adipocytes and the number of adipocytes in the body. Adipocyte number is regulated by the differentiation of cells into adipocytes, and understanding this process is important for identifying novel methods for controlling obesity.Adipocyte differentiation requires expression of the nuclear hormone receptor peroxisome proliferator-activated receptor (PPAR) 2 ␥ (3, 4). When activated by its ligand, PPAR␥ functions as a transcription factor to induce numerous genes involved in adipogenesis. Identification of the endogenous ligand for PPAR␥ has been controversial, but treatment with the prostanoid 15-deoxy-⌬ 12,14 -prostaglandin (PG) J 2 (15d-PGJ 2 ) has long been...

show abstract

“…We also grew tissue explants in collagen gel matrix as described in Sonoda et al (2008) to examine adipokine production. Adipose samples were taken during the hyperphagic season from four 2-year-old male bears (A, B, C, and D).…”

Section: B Differentiated Cellsmentioning

confidence: 99%

A protocol for the isolation and cultivation of brown bear (Ursus arctos) adipocytes

et al. 2016

View full text Add to dashboard Cite

Brown bears (Ursus arctos) exhibit hyperphagia each fall and can become obese in preparation for hibernation. They do this without displaying the physiological problems typically seen in obese humans, such as Type 2 diabetes and heart disease. The study of brown bear hibernation biology could therefore aid in the development of novel methods for combating metabolic diseases. To this end, we isolated mesenchymal stem cells from subcutaneous fat biopsies, and culture methods were developed to differentiate these into the adipogenic lineage. Biopsies were taken from 8 captive male (N = 6) and female (N = 2) brown bears, ages 2-12 years. Plastic adherent, fibroblast-like cells were proliferated and subsequently cryopreserved or differentiated. Differentiation conditions were optimized with respect to fetal bovine serum content and time spent in differentiation medium. Cultures were characterized through immunostaining, RT-qPCR, and Oil red O staining to quantify lipid accumulation. Adiponectin, leptin, and glycerol medium concentrations were also determined over the course of differentiation. The culturing protocol succeeded in generating hormone-sensitive lipase-expressing, lipid-producing white-type adipocytes (UCP1 negative). Serum concentration and time of exposure to differentiation medium were both positively related to lipid production. Cells cultured to low passage numbers retained similar lipid production and expression of lipid markers PLIN2 and FABP4. Ultimately, the protocols described here may be useful to biologists in the field investigating the health of wild bear populations and could potentially increase our understanding of metabolic disorders in humans.

show abstract

A New Organotypic Culture of Adipose Tissue Fragments Maintains Viable Mature Adipocytes for a Long Term, Together with Development of Immature Adipocytes and Mesenchymal Stem Cell-Like Cells

Cited by 37 publications

References 20 publications

Adipose tissue-organotypic culture system as a promising model for studying adipose tissue biology and regeneration

Adipose tissue-organotypic culture system as a promising model for studying adipose tissue biology and regeneration

Cyclooxygenase-2 Deficiency Attenuates Adipose Tissue Differentiation and Inflammation in Mice

A protocol for the isolation and cultivation of brown bear (Ursus arctos) adipocytes

Contact Info

Product

Resources

About