A toolset for 3D in vitro tissue engineering
Tissue-engineered approaches are required to better understand the causes of renal
failure and for the development of new treatment options. Upon renal failure, due to
acute or chronic causes, renal replacement therapies such as dialysis or transplantation
are necessary to restore function. Currently, 185,000 Americans have a functioning
kidney transplant and 450,000 are on dialysis... read more. However, recapitulating the functions of
the human kidney in vitro remains challenging due to the anatomical complexity required
to mimic renal physiology. Despite these challenges, advancements in microfluidic and 3D
tissue culture techniques have demonstrated the importance of both microenvironment and
mechanosensory stimulation in establishing physiologically relevant, in vitro models for
disease studies and drug development. Accordingly, we have developed a 3D in vitro
tissue toolset that can be utilized to achieve the necessary phenotypes for studying
kidney development and disease. In order to understand the development of polycystic
kidney disease we established static hydrogel cultures to characterize long-term
development of cysts and changes in structural morphologies. Additionally, we developed
a modular, three dimensional perfusion culture system to support the controlled fluidic
stimulation of a planar cell layer seeded on a 3D porous, silk protein scaffold. Lastly,
we established a simple, robust assay for the in vitro formation of renal epithelial
tubules by a telomerase immortalized human proximal tubule cell line (RPTEC/tert1). This
methodology yields polarized, luminal tubules that were responsive to TGFβ
stimulation and co-culture with stromal cells. These in vitro model systems, which yield
physiologically relevant phenotypes without complex differentiation protocols or culture
methods, comprise a necessary toolset for future in vitro studies of disease
pathogenesis and nephrotoxicity.
Thesis (Ph.D.)--Tufts University, 2017.
Submitted to the Dept. of Biomedical Engineering.
Advisor: David Kaplan.
Committee: Ron Perrone, Lauren Black, Barbara Ehrlich, and David Kaplan.
Keyword: Biomedical engineering.read less