We have developed a novel, 3D bioreactor technology that permits the long-term culture of adipocytes, which is not possible using traditional 2D cell culture methods. In this study, we will utilize our technology to rapidly and effectively screen the effects of drugs on human adipose tissue function. We will examine the function of adipocytes in both obese and non-obese patients. One of the parameters we will study is cytokine/adipokine expression. With the bioreactor technology, we are able to rapidly and easily analyze daily expression of cytokines in the media. New drugs may target cytokine expression of adipocytes. It has been shown that involved cytokine behavior in obesity includes the increased expression of, but not limited to: MCP-1 (monocyte chemotactic protein-1, which can recruit macrophages to adipose tissue), TNF-1 (tumor necrosis factor-1, a pro-inflammatory mediator secreted by macrophages), and IL-8 (interleukin-8, a pro-inflammatory cytokine secreted by macrophages). Also of interest is the expression of anti-inflammatory cytokines, such as IL-10 (interleukin-10). While these mediators have been examined using human and murine adipose tissue in 2D in vitro culture, improved experimental systems are necessary to allow the development of high throughput assays for drug discovery. Therefore, the specific aims of this proposal are to: 1) Isolate and characterize human adipose-derived stem cells from both male and female patients, age 40-60 years, (non-obese, vs. obese patients);2) Develop a novel, multi-compartment, hollow fiber 3D perfusion bioreactor technology for ASC culture in 3D bioreactor;3) Utilize the 3D perfusion bioreactor system as a tool to study the effects of drug therapies on adipose function. In summary, cell-cell contact in a 3D culture system mimicking natural adipose tissue represents an improvement over current petri-dish technologies aimed at developing high throughput assays for drug discovery.

Public Health Relevance

Nation-wide obesity rates continue to climb at an alarming rate. Different approaches to controlling the obesity epidemic include expanding education programs about healthy living, reducing the marketing of unhealthy, high-fat foods, and providing low-cost exercise programs. As scientists, we can contribute to a reduction in obesity by increasing our understanding of the mechanisms of adipose cell function and the high throughput screening of new drugs. As such, this proposal describes an innovative, engineering approach to studying adipose tissue in a native, 3D architecture, utilizing a novel 3D hollow fiber bioreactor which permits both the long-term 3D culture of adipocytes and the rapid screening of drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK089190-02
Application #
8119423
Study Section
Biomaterials and Biointerfaces Study Section (BMBI)
Program Officer
Haft, Carol R
Project Start
2010-09-01
Project End
2013-08-31
Budget Start
2011-09-01
Budget End
2012-08-31
Support Year
2
Fiscal Year
2011
Total Cost
$306,987
Indirect Cost
Name
University of Pittsburgh
Department
Surgery
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Gerlach, Jorg C; Lin, Yen-Chih; Brayfield, Candace A et al. (2012) Adipogenesis of human adipose-derived stem cells within three-dimensional hollow fiber-based bioreactors. Tissue Eng Part C Methods 18:54-61