In vitro fibrosis model, preparing method therefor, and use thereof
Inventors
Kim, Sang Heon • Park, Kwi Deok • LEE, Kang Won • RAJANGAM, Thanavel
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11365396-B2
Publication Date
2022-06-21
Expiration Date
Abstract
Provided are an in vitro fibrosis model, a method of preparing the in vitro model, and use of the in vitro model, the in vitro model including a cell cluster differentiated from mesenchymal cells, wherein the cell cluster exhibits pathological characteristics of fibrosis.
Core Innovation
The invention describes a method of screening a candidate therapeutic agent for treatment of a fibrotic disease by culturing adipose stem cells to form a three-dimensional (3D) cell cluster. The adipose stem cells adhere to a culture container comprising a hydrophobic surface, and the hydrophobic surface is coated with a growth factor immobilized to the hydrophobic surface through a polypeptide linker.
The growth factor comprises vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), platelet-derived endothelial growth factor (PDGF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), or a heparin-binding domain (HBD). The polypeptide linker comprises a maltose-binding protein (MBP), a hydrophobin, or a hydrophobic cell penetrating peptide (CPP).
The 3D cell cluster is cultured to provide an in vitro fibrosis model comprising the 3D cell cluster. The in vitro fibrosis model is treated with a test substance, and a test substance is selected as a candidate therapeutic agent if it exhibits improvement or treatment of pathological characteristics of fibrosis in the 3D cell cluster or cells thereof as compared with an untreated control group.
Claims Coverage
The independent claim includes 3 inventive features covering a 3D cell cluster-based in vitro fibrosis model and screening selection. Dependent claims further define the cluster geometry and size, fibrosis readouts, and the scope of fibrotic disease.
Hydrophobic surface with growth factor immobilized via polypeptide linker for forming 3D cell clusters
Culturing adipose stem cells to form a three-dimensional (3D) cell cluster by adhering the adipose stem cells to a culture container comprising a hydrophobic surface, wherein the hydrophobic surface is coated with a growth factor immobilized to the hydrophobic surface through a polypeptide linker.
In vitro fibrosis model from culturing the 3D cell cluster
Culturing the 3D cell cluster to provide an in vitro fibrosis model comprising the 3D cell cluster.
Treating with test substance and selecting based on improved fibrosis pathological characteristics
Treating the in vitro fibrosis model comprising the 3D cell cluster with a test substance, and selecting a test substance as a candidate therapeutic agent for treatment of the fibrotic disease if it exhibits improvement or treatment of pathological characteristics of fibrosis in the 3D cell cluster or cells thereof as compared with an untreated control group.
Spherical 3D cell cluster with defined diameter range
The 3D cell cluster is spherical with a diameter in a range of about 300 μm to about 2,000 μm.
Fibrosis characterized by collagen and fibrosis markers
Fibrosis is characterized by increased collagen deposition, increased expression, secretion, or synthesis of TGF-beta, Smad, laminins, and smooth muscle actin (SMA), and increased collagen-induced cell death in the 3D cluster compared with a two-dimensional culture of adipose stem cells.
Fibrotic disease includes listed fibrotic disorders
The fibrotic disease includes at least one selected from idiopathic pulmonary fibrosis (IPF), pulmonary fibrosis, interstitial lung disease, cardiac, renal, liver, and skin fibrotic disorders, and additional specified fibrosis-related conditions.
The claims center on generating an in vitro fibrosis model by culturing adipose stem cells into 3D clusters on a hydrophobic culture container with growth factor immobilized via a polypeptide linker, then treating the model with a test substance and selecting candidates based on improvement of fibrosis pathological characteristics versus an untreated control. Dependent claims add the spherical cluster diameter range, fibrosis-related readouts, and an enumerated scope of fibrotic diseases.
Stated Advantages
Documented Applications
No documented applications found
Interested in licensing this patent?