A study of adipocyte differentiation of 3T3-L1 cells in LbL nano film coated 3D microbead based microenvironment

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The following specific targets have been accomplished in this thesis work: generation of polymer microbeads using droplet microfluidics, generation of LbL nanofilm library using CF-LbL assembly technique, characterization of nanofilm library, culturing 3T3-L1 cells on different LbL nanofilms to observe cell viability, selection of suitable film for cell culture, modifying microbeads with suitable LbL nanofilms, culturing 3T3-L1 on surface modified microbead-based 3D microenvironment and observation of adipocyte differentiation. Specifically, adipocyte differentiation was observed on a plane surface, hydrogel surface and in microbead-based 3d microenvironment for both with and without surface modification with the LbL nanofilms, under the condition with or without the presence of an insulin inducer. Selection of LbL nanofilm films was done by observing the survival of the cell on different polyelectrolyte films using live/dead cell viability assay. The films were studied in terms of their thickness, degradation profile in presence of activated enzymes. Study on aggregation of microbeads was used to determine charge density of the polyelectrolyte films. Alginate microbeads was prepared by droplet microfluidics and the gelation of microbeads was based on physical crosslinking of alginate with ca+2 ions, respectively. Surface morphology and internal polymer network structure of alginate micro-gel beads were studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), respectively. Water retention and swelling ratio of the alginate hydrogels were also determined.

Adipocyte differentiation, 3T3-L1, Alginate micro-bead, Three-dimensional (3-D) microenvironment, Layer-by-layer (LbL)