Well plate-based perfusion culture model of endosteal-extracellular matrix (ecm)-and endothelial-myeloma interactions and methods for testing personalized therapeutics for multiple myeloma

An ex vivo model of a three dimensional (3D) cellular network found in native bones via biomimetic assembly of osteocytes and microbeads in a microfluidic perfusion culture device comprising

• (a) preparing an in vitro multiwell plate-based perfusion culture device, comprising, from top to bottom:
• 1. a bottomless multi-well plate comprising a plurality of bottomless wells;
• 2. a first micropatterned polymer layer attached to a bottom surface of the bottomless multi-well plate to form a plurality of adjacent wells, one or more of each pair of adjacent wells comprising a transparent polymer membrane placed within the one or more of each pair of adjacent wells;
• 3. a second micropatterned polymer layer comprising two or more holes that correspond to two or more adjacent wells, the second micropatterned polymer layer being attached to a bottom surface of the first micropatterned polymer layer, such that each hole of the second micropatterned polymer layer is aligned with the two or more adjacent wells in the first micropatterned polymer layer, one or more of each pair of adjacent wells comprising the transparent polymer membrane;
• 4. a microfluidic channel formed between the two adjacent wells that allows internal fluidic communication between the two adjacent wells;
• 5. one or more removable polymer plugs, each located at a top surface of each of the plurality of wells, and one or more tubes, each connected to the one or more polymer plugs;
• 6. a pump connected to a reservoir that removably connects to the tubes;
• 7. a transparent, optical grade glass layer attached to the bottom surface of the second micropatterned polymer layer that forms a bottom surface for the plurality of wells and that seals the multi-well plate perfusion culture device;
• wherein
• (i) one or more of the two adjacent wells is a cell culture chamber comprising a first well region including a first well and a second well region including a second well;
• (ii) the microfluidic channel connects the first well region and the second well region with one another;
• (iii) the first well is adapted to receive a therapeutic agent, the second well is adapted to receive a biological sample of cells;
• and
• (iv) liquids, nutrients and dissolved gas molecules flow through the channel
• (b) constructing an ex vivo endosteal microenvironment perfused by nutrients and dissolved gas molecules by;
• 1. seeding a surface of the culture chamber of the device of (a) with
• (i) microbeads;
• (ii) osteocyte cells (OSTs);
• and
• (iii) osteoblast cells (OSBs), and
• 2. culturing the cells with a culture medium through the microfluidic channel for a time effective for the cells to form three-dimensional (3D) nodular structures that comprise a 3D-endosteal-like tissue.