Recombinant Human Connective Tissue Growth Factor

Recombinant Human Connective Tissue Growth Factor (rhCTGF) is widely used in research due to its critical roles in cell proliferation, differentiation, extracellular matrix (ECM) production, angiogenesis, and tissue repair, making it highly valuable for studies in regenerative medicine, fibrosis, wound healing, and tissue engineering.

Key reasons to use rhCTGF in research applications:

• Regenerative Medicine and Tissue Engineering: rhCTGF is a potent regulator of tissue growth and repair. It promotes fibroblast proliferation, ECM synthesis (including collagen and fibronectin), and angiogenesis, all of which are essential for tissue regeneration and wound healing. For example, rhCTGF accelerates wound closure and increases tissue stiffness by enhancing collagen IV deposition and smooth muscle actin expression in wound models.

• Fibrosis and Disease Modeling: CTGF is a central mediator in fibrotic diseases (e.g., pulmonary, liver, and kidney fibrosis) and cancer, making it a valuable tool for modeling these conditions and testing anti-fibrotic therapies. Its ability to interact with TGF-β and other signaling pathways allows researchers to dissect mechanisms of fibrosis and tissue remodeling.

• Bone and Cartilage Research: rhCTGF stimulates osteoblast differentiation and bone formation, supporting its use in bone regeneration and repair studies. It also promotes chondrocyte proliferation and proteoglycan synthesis, making it relevant for cartilage tissue engineering.

• Angiogenesis and Cell Communication: CTGF induces angiogenesis and enhances cell-cell communication via pathways such as PI3K/Akt and integrin signaling, which are crucial for vascular biology and tissue integration studies.

• Experimental Consistency and Reproducibility: Recombinant growth factors like rhCTGF offer high purity, batch-to-batch consistency, and superior bioactivity compared to native proteins, ensuring reproducible results in cell culture, organoid, and in vivo models.

• Reduction of Animal-Derived Variability: Using recombinant proteins minimizes the risk of pathogen transmission and immunogenicity associated with animal-derived materials, supporting safer and more ethical research practices.

Applications include:

• Wound healing and chronic wound models
• Fibrosis and anti-fibrotic drug screening
• Bone and cartilage regeneration
• Angiogenesis assays
• Organoid and stem cell differentiation protocols

In summary, rhCTGF is a versatile and reliable tool for investigating mechanisms of tissue repair, fibrosis, and regeneration, and for developing new therapeutic strategies in these areas.

Yes, Recombinant Human Connective Tissue Growth Factor (CTGF) can generally be used as a standard for quantification or calibration in ELISA assays, provided that:

1. The recombinant protein is biologically active and structurally similar to the endogenous CTGF present in your samples. Most commercially available recombinant CTGF is designed to mimic the native protein and is suitable for this purpose.

2. The ELISA kit you are using is compatible with the recombinant standard. Most sandwich ELISA kits for human CTGF are validated to detect both natural and recombinant forms of the protein, as long as the epitopes recognized by the capture and detection antibodies are present on the recombinant molecule.

3. You prepare the standard curve appropriately. Serial dilutions of the recombinant CTGF should be made in the same matrix as your samples (e.g., assay buffer, serum, or plasma) to minimize matrix effects and ensure accurate quantification.

4. The concentration range of your standard curve covers the expected levels in your samples. If the recombinant CTGF is supplied at a high concentration, you may need to dilute it to fall within the detection range of your ELISA kit.

Best Practices:

• Always check the datasheet or product information for both the recombinant CTGF and the ELISA kit to confirm compatibility.
• Run the standard curve in duplicate or triplicate for each assay.
• If possible, validate the use of the recombinant standard by spiking known amounts into your sample matrix and assessing recovery.

In summary, recombinant human CTGF is commonly used as a standard in ELISA assays for CTGF quantification, but compatibility and proper preparation are essential for accurate results.

Recombinant Human Connective Tissue Growth Factor (CTGF/CCN2) has been validated in published research for applications including wound healing, fibrosis modeling, cell differentiation, angiogenesis, tissue engineering, and immune modulation.

Key validated applications in the literature include:

• Wound Healing Models: Recombinant CTGF has been topically applied to diabetic rodent wounds, resulting in accelerated wound closure, increased granulation tissue, enhanced collagen IV accumulation, and improved mechanical strength of healed skin. These effects have also been correlated with endogenous CTGF levels in human diabetic foot ulcers, supporting its functional role in human wound healing.

• Fibrosis and Fibrotic Disease Models: CTGF is widely used to study fibrogenesis, particularly in the context of TGF-β-driven fibroblast activation, extracellular matrix (ECM) production, and tissue remodeling. Recombinant CTGF is applied in vitro to induce or enhance fibrotic responses in fibroblasts and other cell types, and in vivo to model fibrotic diseases of the lung, liver, kidney, and heart.

• Cell Proliferation, Differentiation, and EMT: CTGF has been used to stimulate proliferation and differentiation of various cell types, including fibroblasts and granulosa cells. It is also validated for inducing epithelial-mesenchymal transition (EMT)-like changes in epithelial cells, relevant for cancer and fibrosis research.

• Angiogenesis and Tissue Engineering: Recombinant CTGF is incorporated into bioactive scaffolds and biomaterials to promote both angiogenesis (formation of new blood vessels) and osteogenesis (bone formation) in tissue engineering and regenerative medicine applications.

• Immunomodulation: The C-terminal module of CTGF has been shown to regulate Th17 cell differentiation in vitro, implicating recombinant CTGF in studies of immune cell polarization and chronic inflammation.

• Signaling Pathway Studies: Recombinant CTGF is used to dissect signaling pathways, particularly its interactions with TGF-β, Smad, and ERK pathways, in various cell types.

Experimental formats validated:

• In vitro cell culture (e.g., fibroblasts, granulosa cells, T cells)
• In vivo animal models (e.g., rodent wound healing, fibrosis models)
• Bioengineering approaches (e.g., incorporation into scaffolds, microencapsulation)

Assays and endpoints:

• Cell proliferation and migration assays
• ECM protein expression (e.g., collagen, fibronectin)
• Wound closure and tissue strength measurements
• Flow cytometry for immune cell phenotyping
• Histological and immunohistochemical analyses

These applications are supported by multiple peer-reviewed studies, demonstrating the versatility of recombinant human CTGF as a research tool in cell biology, disease modeling, regenerative medicine, and immunology.

To reconstitute and prepare Recombinant Human Connective Tissue Growth Factor (CTGF) for cell culture experiments, follow these best-practice steps:

• Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
• Add sterile, distilled water or the recommended buffer to achieve a concentration between 0.1–1.0 mg/mL. Some protocols specify at least 100 μg/mL as a minimum concentration.
• Do not vortex; instead, gently pipette the solution up and down or swirl to dissolve the protein completely.
• Allow the solution to sit at room temperature for several minutes to ensure full dissolution.
• If the product datasheet specifies a particular buffer (e.g., PBS or 5 mM sodium acetate, pH 6.0), use that instead of water. Always check the product-specific instructions.
• For cell culture applications, after initial reconstitution, further dilute the protein in cell culture medium or buffer containing a carrier protein such as 0.1% BSA to stabilize the protein and minimize adsorption to plasticware.
• Aliquot the solution to avoid repeated freeze-thaw cycles. Store aliquots at –20°C to –80°C for long-term storage, or at 2–8°C for up to one week.
• Avoid multiple freeze/thaw cycles to preserve protein activity.

Summary Table: Reconstitution and Preparation

Additional notes:

• Always consult the specific product datasheet for any unique requirements, as some preparations may require a particular buffer or pH.
• For cell-based assays, dilute the reconstituted CTGF to the desired working concentration in cell culture medium just before use.
• If using for ELISA or other biochemical assays, follow the assay-specific dilution and buffer recommendations.

These steps will help ensure protein stability and biological activity for your cell culture experiments.