Compound Distribution Kit Protocol

Introduction

In both in vitro and in vivo experiments, researchers must consider compound distribution within the biological model and experimental setup prior to quantitative drug studies, as distribution determines exposure — the concentration of a compound that cells truly experience.

In in vivo systems, this is addressed by volume of distribution studies, which relate compound dosage to its effective concentration. However, in both in vivo and in vitro studies, the distribution effects of system components, such as infusion tubing, syringes, tissue-culture plates, and pipette tips, are often missed.

With Organ-Chip experiments, we proactively address compound distribution in a number of ways. Several of these are embedded in our protocol designs, where we have selected experimental conditions to optimize compound exposure. Additionally, we have developed the Compound Distribution Kit to directly evaluate distribution and compound exposure.

The Compound Distribution Kit is intended to be used as a specialized control experiment — the distribution control experiment — prior to the intended Organ-Chip study. As such, the contents of the Compound Distribution Kit mirror the contents of the Organ-Chip Bio-Kit, and the protocol used for the distribution control experiment mirrors a simplified version of the intended study (e.g., without cells or ECM coating). The distribution control experiment’s output indicates whether any compound may be distributed into the system and away from cells. Moreover, in some cases, the distribution control can be used to quantitatively correct the experimental results of the intended study and assign it appropriate error bars.

Immunofluorescence Staining of Stabilin-2

Introduction

This protocol instructs users how to visualize the multifunctional scavenger receptor Stabilin-2 in Emulate Organ-Chips via immunofluorescence.

Stabilin-2 is a large, transmembrane receptor protein and is primarily expressed on sinusoidal endothelial cells of liver, spleen, and lymph node. The receptor has been shown to endocytose ligands such as low density lipoprotein, Gram-positive and Gram-negative bacteria, and advanced glycosylation end products.

Liver-Chip Culture Protocol // Quad-Culture

Introduction

(As of August 14, 2023, a new version of the protocol has been uploaded.)

This protocol covers how to seed and culture a Quad-Culture Liver-Chip using the Chip-S1™ stretchable chip.

Liver-Chip Culture Protocol // Co-Culture

(As of August 14, 2023, a new version of the protocol has been uploaded.)

Introduction

This protocol covers how to seed and culture a Co-Culture Liver-Chip using the Chip-S1® Stretchable Chip.

Cell Lysis for RNA Isolation

Introduction

Goal: Lyse cells in Emulate Organ-Chips for RNA isolation.