Methods of tissue generation and tissue engineered compositions
View Patent ↗Provided are methods and compositions for constructing stable mammalian embryonic epithelial tissues and organs as well as constructing kidney tissue, and treating renal failure. Disclosed are methods of using an active epithelial growth factor having the capability of effectuating induction of growth and morphogenesis is cells.
1. A method of generating a tubular tissue structure, comprising:
(a) contacting a Wolffian duct (WD) cell with GDNF and FGF1 to stimulate growth and proliferation;
(b) contacting the cells with PTN and GDNF to promote formation of tubular tissue branches and/or globular morphology to generate a ureteric bud;
(c) combining the ureteric bud tissue with metanephric mesenchyme in a biocompatible matrix; and
(d) culturing the combination to form a tubular tissue or kidney tissue in vitro.
2. A method for in vitro culturing and propagating Wolffian duct bud tissue, comprising:
isolating Wolffian duct bud tissue from mesenchyme tissue obtained from embryonic kidney rudiments;
culturing the isolated Wolffian duct bud tissue in a biocompatible matrix in the presence of a culture medium comprising FGF1 and GDNF to produce tubular branches within the biocompatible matrix;
separating the plurality of branch tips to generate bud fragments; and
culturing each of the bud fragments in a biocompatible matrix with a culture medium comprising pleiotrophin.