Engineering and Characterization of a Biomimetic Microchip for Differentiating Mouse Adipocytes in a 3D Microenvironment

Pharm Res. 2022 Feb;39(2):329-340. doi: 10.1007/s11095-022-03195-0. Epub 2022 Feb 15.

Abstract

Although two-dimensional (2D) cell cultures are the standard in cell research, one pivotal disadvantage is the lack of cell-cell and cell-extracellular matrix (ECM) signaling in the culture milieu. However, such signals occur in three-dimensional (3D) in vivo environments and are essential for cell differentiation, proliferation, and a range of cellular functions. In this study, we developed a microfluidic device to proliferate and differentiate functional adipose tissue and adipocytes by utilizing 3D cell culture technology. This device was used to generate a tissue-specific 3D microenvironment to differentiate 3T3-L1 preadipocytes into either visceral white adipocytes using visceral adipose tissue (VAT) or subcutaneous white adipose tissue (SAT). The microchip has been tested and validated by functional assessments including cell morphology, inflammatory response to a lipopolysaccharide (LPS) challenge, GLUT4 tracking, and gene expression analyses. The biomimetic microfluidic chip is expected to mimic functional adipose tissues that can replace 2D cell cultures and allow for more accurate analysis of adipose tissue physiology.

Keywords: 3D Cell Culture; Adipocyte differentiation; Adipocytes; Adipose tissue; Microfluidics; Organ-on-a-Chip.

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes, White / drug effects
  • Adipocytes, White / metabolism
  • Adipocytes, White / physiology*
  • Adipogenesis*
  • Animals
  • Biomimetic Materials*
  • Cell Culture Techniques, Three Dimensional / instrumentation*
  • Cell Proliferation
  • Cyclooxygenase 2 / genetics
  • Cyclooxygenase 2 / metabolism
  • Cytokines / genetics
  • Cytokines / metabolism
  • Female
  • Glucose Transporter Type 4 / metabolism
  • Inflammation Mediators / metabolism
  • Lab-On-A-Chip Devices*
  • Lipopolysaccharides / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Microfluidic Analytical Techniques / instrumentation*

Substances

  • Cytokines
  • Glucose Transporter Type 4
  • Inflammation Mediators
  • Lipopolysaccharides
  • Slc2a4 protein, mouse
  • Ptgs2 protein, mouse
  • Cyclooxygenase 2