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An Open-Source Normothermic Perfusion System Designed for Research Scientists
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Long term perfusion system supporting adipogenesis.

Rosalyn D Abbott1, Waseem K Raja2, Rebecca Y Wang1

  • 1Department of Biomedical Engineering, Science and Technology Center, Tufts University, 4 Colby St, Medford, MA 02155, United States.

Methods (San Diego, Calif.)
|April 7, 2015
PubMed
Summary
This summary is machine-generated.

Three-dimensional (3D) perfusion culture using silk biomaterials creates robust engineered adipose tissue models. This method enhances adipogenesis and provides a sustainable platform for studying diseases and regenerative medicine.

Keywords:
Adipose tissue engineeringLong term culturePerfusionSilk scaffoldThree dimensional culture

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Cell Biology

Background:

  • Adipose tissue models are crucial for understanding diseases and developing regenerative strategies.
  • Perfusion systems offer more physiologically relevant adipose tissue models but pose culturing challenges for adipocytes.
  • Existing methods require improved techniques for long-term in vitro adipogenic cultures.

Purpose of the Study:

  • To develop and evaluate two perfusion culture systems (2D and 3D) for long-term in vitro adipogenic cultures.
  • To test the hypothesis that a silk protein biomaterial scaffold in a 3D perfusion system would yield a more sustainable and physiologically relevant adipose tissue model than 2D culture.
  • To compare the efficacy of 2D and 3D perfusion systems in supporting adipogenesis and adipocyte viability.

Main Methods:

  • Development of two perfusion culture systems: a 2D system and a 3D system utilizing a silk protein biomaterial scaffold.
  • Culturing adipocytes within these perfusion systems for extended periods to assess adipogenesis.
  • Quantification of adipogenesis, lipogenesis, lipolysis, glycerol secretion, and lactate dehydrogenase (LDH) release as key performance indicators.

Main Results:

  • Both 2D and 3D perfusion systems supported adipogenesis, but the 3D system demonstrated superior robustness.
  • The 3D silk scaffold provided essential mechanical structure, preventing the loss of fragile adipocytes observed in 2D cultures.
  • 3D perfusion cultures exhibited enhanced lipogenesis and lipolysis, reduced LDH secretion, and increased glycerol secretion compared to 2D cultures.

Conclusions:

  • 3D perfusion culture with silk biomaterial scaffolds is a robust method for generating sustainable adipose tissue models.
  • This approach overcomes challenges associated with culturing fragile adipocytes in perfusion systems.
  • The developed 3D system holds significant promise for long-term in vitro studies of disease mechanisms and for soft tissue regenerative medicine applications.