Related Experiment Video

Updated: Aug 4, 2026

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers
18:57

Models and Methods to Evaluate Transport of Drug Delivery Systems Across Cellular Barriers

Published on: October 18, 2013

Perforated MDCK cells support intracellular transport

K Simons1, H Virta

  • 1European Molecular Biology Laboratory, Heidelberg, FRG.

The EMBO Journal
|August 1, 1987
PubMed

Insights

Researchers created perforated Madin-Darby canine kidney (MDCK) cells that retain function for studying intracellular transport. This method allows macromolecule entry, enabling efficient analysis of protein and lipid trafficking pathways.

Area of Science:

  • Cell Biology
  • Membrane Biology
  • Molecular Biology

Background:

  • Intracellular transport is crucial for cellular function.
  • Studying transport mechanisms often requires cell-free systems, which can be complex.
  • A need exists for methods that allow direct manipulation of cellular contents while maintaining cell viability.

Purpose of the Study:

  • To develop a novel method for perforating the plasma membrane of MDCK cells.
  • To assess the functional integrity of perforated cells.
  • To demonstrate the utility of perforated cells for studying intracellular transport.

Main Methods:

  • Applying a nitrocellulose acetate filter to the apical surface of MDCK cells grown on coverslips.
  • Drying the filter to adhere membrane segments, then peeling it off to create perforations.
  • Utilizing light and electron microscopy to assess cell integrity.
  • Tracking protein and lipid transport using radiolabeling and fluorescent probes.

Main Results:

  • Perforation of the plasma membrane was achieved while maintaining cell structure and function.
  • Perforated cells successfully supported vesicular transport of proteins (G protein) and lipids (sphingolipids).
  • Newly synthesized proteins were correctly processed and trafficked to the trans Golgi.
  • Lipid probes showed transport from the Golgi to the basolateral surface.

Conclusions:

  • The developed method effectively perforates MDCK cell plasma membranes, allowing macromolecule entry.
  • Perforated cells remain functional and are suitable for studying intracellular transport mechanisms.
  • This technique offers a convenient and efficient alternative to cell-free assays for investigating molecular mechanisms of intracellular trafficking.

More Related Videos

A Choroid Plexus Epithelial Cell-based Model of the Human Blood-Cerebrospinal Fluid Barrier to Study Bacterial Infection from the Basolateral Side09:58

A Choroid Plexus Epithelial Cell-based Model of the Human Blood-Cerebrospinal Fluid Barrier to Study Bacterial Infection from the Basolateral Side

Published on: May 6, 2016

An Intravital Microscopy-Based Approach to Assess Intestinal Permeability and Epithelial Cell Shedding Performance07:32

An Intravital Microscopy-Based Approach to Assess Intestinal Permeability and Epithelial Cell Shedding Performance

Published on: December 3, 2020

Related Concept Videos

Pinocytosis00:43

Pinocytosis

Cells use energy-requiring bulk transport mechanisms to transfer large particles, or large amounts of small particles, into or out of the cell. The cells envelop the particles in spherical membranes called vesicles or vacuoles. Vesicles that transport material into the cell are built from the cell membrane. These vesicles encapsulate external molecules and transport them into the cell in a process called endocytosis.
Transcellular Transport of Solutes01:23

Transcellular Transport of Solutes

Transcellular transport of solutes is the movement of substances like monosaccharides and amino acids through polarized cells. This transport mechanism is primarily seen in epithelial and endothelial cells aided by membrane transport proteins such as channels and transporters. The tight junctions between these cells confine the membrane proteins to the two sides of the cell. The epithelial cells have distinct apical and basolateral domains. In contrast, the endothelial cells show the luminal...
Protein Transport into the Inner Mitochondrial Membrane01:34

Protein Transport into the Inner Mitochondrial Membrane

Nuclear encoded mitochondrial precursors are imported to the inner membrane in a multistep process involving two separate translocons, TIM22 and TIM23. TIM23 is a cation-selective pore that remains closed by the N terminal segment of the protein. Negative charges on the TIM23 act as a receptor for the incoming precursor, pulling the positively charged matrix-targeting sequence for peptide insertion and translocation.
Transport of mitochondrial precursors across the TIM23 channel is driven by...
Introduction to Membrane Traffic01:44

Introduction to Membrane Traffic

The ER, Golgi apparatus, endosomes, and lysosomes work in tandem to modify, sort, and package proteins and lipids. An integrated membrane trafficking network facilitates the back and forth shuttling of molecules within different organelles in the same cell or across the cell membrane.
The transport of soluble and membrane proteins is mediated by transport vesicles that collect cargo from one cellular compartment and deliver it to another by fusing with the target organelle membrane. The Rab...
Cellular Membranes and Drug Transport01:24

Cellular Membranes and Drug Transport

Drugs must traverse multiple biological barriers, such as multi-layered skin, single-layered intestinal epithelium, and the plasma membrane, to reach their target sites within the body. The plasma membrane, a highly structured composite of phospholipids, carbohydrates, and proteins, is the cell's protective boundary, facilitating selective substance exchange.
Phospholipids arrange themselves into a bilayer, with hydrophilic heads oriented outward and hydrophobic tails facing inward.
Facilitated Diffusion01:16

Facilitated Diffusion

The plasma membrane, a critical structure in cellular biology, houses an array of transporters, or carrier proteins, interspersed within its lipid bilayer. These proteins play a crucial role in solute transport through facilitated diffusion, a form of passive diffusion that uses transporters to move the molecules across the membrane.
In this process, substrates such as organic compounds and ions interact with a transporter on one side, triggering conformational changes in proteins that enable...