Related Experiment Video
Updated: Jan 26, 2026
09:05
In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition
Published on: August 20, 2016
8.4K
3D in vitro model for human corneal endothelial cell maturation
Audrey E K Hutcheon1, James D Zieske1, Xiaoqing Guo1
1Schepens Eye Research Institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, 20 Staniford Street, Boston, MA, 02114, USA.
Experimental Eye Research
|April 14, 2019
Summary
A novel 3D co-culture model using human corneal fibroblasts (HCF) and human corneal endothelial cells (HCEndoCs) promotes in vivo-like endothelial maturation and new Descemet
Area of Science:
- Ophthalmology
- Regenerative Medicine
- Cell Biology
Background:
- The corneal endothelium is crucial for maintaining corneal clarity.
- Current in vitro models do not fully replicate the in vivo environment for endothelial cell studies.
- Understanding endothelial cell behavior is vital for developing new therapies like corneal endothelial cell injection.
Purpose of the Study:
- To assess the feasibility of a 3D co-culture model using human corneal fibroblasts (HCF) and human corneal endothelial cells (HCEndoCs).
- To observe the development and maturation of HCEndoCs in this 3D stromal construct.
- To evaluate the potential of this model for studying corneal endothelial cell injection therapies.
Main Methods:
- Developed a 3D HCF construct using HCFs and their self-assembled matrix.
- Co-cultured primary (pHCEndoC) and cell line (HCEndoCL) HCEndoCs on the 3D constructs.
- Utilized light microscopy, transmission electron microscopy (TEM), and indirect-immunofluorescence microscopy (IF) to analyze cell morphology, ultrastructure, tight junction formation (ZO-1), maturation (ALDH1A1), basement membrane formation (Laminin), proliferation (Ki67), cell death (caspase-3), and fibrotic response (CTGF).
Main Results:
- HCEndoCLs on 3D constructs exhibited in vivo-like morphology (uniform, closely packed).
- Co-culture promoted the formation of a Descemet's membrane (DM)-like structure and stimulated laminin localization.
- ALDH1A1 expression indicated HCEndoC maturation, with minimal cell death or proliferation observed; CTGF indicated a fibrotic response.
Conclusions:
- The 3D co-culture model effectively supports HCEndoC maturation and the formation of a DM-like matrix in vitro.
- This model shows promise for investigating corneal endothelial cell injection therapies and understanding the implications of Descemet's membrane removal.
- The model activates a fibrotic response, highlighting the importance of preserving native DM during treatments.
Related Concept Videos
01:28Maturation of Endosomes
5.7K
The early endosome containing internalized molecules matures through transformations in its location, morphology, intraluminal pH, and membrane protein composition. Together, these changes result in a more acidic late endosome that contains multiple intraluminal vesicles; therefore, the late endosome is also called a multivesicular body (MVB).
Changes in location
The maturing endosome moves along microtubules from the periphery of the cell towards the perinuclear region. This movement of the...
Changes in location
The maturing endosome moves along microtubules from the periphery of the cell towards the perinuclear region. This movement of the...
5.7K
01:26Bacterial Protein Maturation
487
Bacterial protein maturation is a tightly regulated process that ensures newly synthesized polypeptides achieve correct functional conformations. This maturation involves a series of modifications, folding events, and quality control steps, often assisted by specialized chaperone proteins.N-Terminal ModificationsThe maturation of bacterial polypeptides begins cotranslationally as the polypeptide exits the ribosome. The first amino acid, N-formylmethionine (fMet), is typically modified at the...
487
01:13In Vitro Drug Dissolution: Compendial Testing Models I
252
Compendial dissolution methods are standardized procedures defined by pharmacopeias to evaluate the rate at which a drug dissolves in a specific medium. These methods ensure batch-to-batch consistency, enable quality control, and support the prediction of drug bioavailability. They are critical for both immediate and modified-release drug products.The apparatuses used for dissolution testing differ in their design and mechanical function, but all aim to simulate the physiological environment of...
252
01:09In Vitro Drug Dissolution: Compendial Testing Models II
290
Various dissolution methods are utilized to assess a drug’s dissolution rate, including the flow-through cell, paddle-over-disk, cylinder, and reciprocating disk methods.The flow-through cell apparatus (USP (United States Pharmacopeia) method 4) comprises a reservoir for the dissolution medium and a pump that propels the medium through the cell containing the test sample. This method is crucial for assessing modified-release dosage forms with minimally soluble active ingredients,...
290
01:16In-vitro Mutagenesis
16.2K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
16.2K
01:24In Vitro Fertilization
926
In vitro fertilization (IVF) is a form of assisted reproductive technology where an egg is fertilized with sperm in a controlled laboratory environment before transferring the resulting embryo into the uterus. This process is designed to help individuals and couples experiencing difficulties conceiving.
The IVF process begins with ovarian stimulation, during which reproductive endocrinologists prescribe hormonal medications to stimulate the ovaries to produce multiple eggs instead of the single...
The IVF process begins with ovarian stimulation, during which reproductive endocrinologists prescribe hormonal medications to stimulate the ovaries to produce multiple eggs instead of the single...
926