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Introduction:

Cell culturing has been around for decades providing a relatively straightforward, affordable, and controlled way to study cell functions and mechanisms, however effects and phenomena seen in  in vitro experiments may not always translate into effects of phenomena seen in vivo.  At ScienCell where we specialize in primary cell cultures, we understand the indispensable utility of in vitro research and have been developing in vitro systems that provide more in vivo relevance using 3-dimensional collagen I matrices.  The first set in this line of new products will focus on mimicking angiogenesis:

3D Tubule Formation Assay Kit: An inclusive kit that uses purified collagen I to mimic all the intricacies of angiogenesis more closely, including the critical lumen formation step.  This kit includes all components necessary to form a 3-dimensional collagen gel dot with endothelial cells embedded within it.  The embedded cells survive, proliferate, migrate, adhere, anastomose, and within 1 week form multicellular, lumen-containing tubular structures.  Embedded gel dots can be fixed and stained at the assay’s termination to aid in quantifying tubule length, area, number, and lumen formation using microscopy.  Fixed gel dots can also be sectioned for more in-depth analysis.

 

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(Top left) Schematic of one gel dot with endothelial cells embedded within it.  Gel dots are cultured in wells of a 24-well plate.  (Top right) Light microscope image of tubules formed from day 5 of embedding.  (Lower right) Higher magnification of a tubule formed from day 5 of embedding under light microscopy.  The green arrow points to a primitive lumen within the tubule.  (Lower left) Immunofluorescent, high-magnification image of fixed tubules from day 6 of embedding.  Endothelial marker CD31 is stained in green (FITC) while nuclei stained with DAPI are represented in blue indicating that tubules are multicellular structures. 

3D Network Formation Assay Kit: An inclusive kit that uses purified collagen I to create a 3-dimensional environment for endothelial network formation.  This kit includes all components necessary to form an endothelial network between a 3-dimensional collagen sandwich.  The sandwiched endothelial cells survive, migrate, adhere, and anastomose in roughly 4 days to enable research on vessel morphogenesis and branching behavior.  Cells can be stained at the assay’s termination to aid in quantifying network area and branch point numbers using microscopy.

 

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(Left) Schematic of one gel sandwich with endothelial cells in it.  Endothelial sandwiches are cultured in wells of a 24-well plate.  (Right) Representative light microscope image of endothelial networks formed from day 4 of a gel sandwich. 

3D Gelling Kit: Utilizes purified collagen I to create an adjustable matrix for culturing endothelial cells.  This kit includes all components necessary to form collagen I matrix of variable thickness and density for culturing endothelial cells.  Collagens are a major structural component in the extracellular matrix and are the primary determinant of extracellular matrix tensile strength.

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Endothelial cells cultured as a monolayer on purified rat-tail collagen I

 

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