Recombinant Human HGF

Recombinant human hepatocyte growth factor (HGF) offers substantial advantages for research applications across multiple disciplines, particularly in regenerative medicine, cell biology, and therapeutic development.

Biological Activity and Consistency

Recombinant human HGF provides high bioactivity and batch-to-batch consistency, which are critical for reproducible research outcomes. This standardization is essential when conducting comparative studies or validating experimental protocols across multiple experiments and timepoints.

Tissue Regeneration and Repair

HGF demonstrates remarkable regenerative potential across diverse tissue types. The protein has shown positive effects in regenerating critical organs including the brain, heart, and liver. Specific applications include:

Neurological applications: HGF administration following transient middle cerebral artery occlusion significantly reduces infarct size and promotes neuronal differentiation. When combined with mesenchymal stem cells (MSCs), HGF enhances motor function recovery and neurological outcomes in intracerebral hemorrhage models.

Spinal cord injury: MSCs modified with HGF demonstrate superior therapeutic outcomes compared to unmodified MSCs in spinal cord ischemia models, with significantly better preservation of motor neurons and lumbar spinal cord integrity.

Metabolic applications: When combined with vascular endothelial growth factor (VEGF), HGF enables transplanted islets to restore normoglycemia in diabetic models, with synergistic effects on vascularization.

Cellular and Developmental Studies

HGF regulates critical cellular processes including epithelial morphogenesis, cell scattering, branching tubulogenesis, and epithelial-mesenchymal transition. These properties make it invaluable for organoid research, stem cell differentiation studies, and functional assays examining cell migration and proliferation.

Clinical Translation

Recombinant HGF has progressed to clinical evaluation, with phase-I and phase-I/II trials investigating intrathecal administration for amyotrophic lateral sclerosis and spinal cord injury. Early clinical studies have also examined HGF efficacy in treating chronic leg ulcers, establishing a foundation for translational research.

Biomarker Applications

Beyond direct therapeutic use, HGF serves as a diagnostic biomarker for various pathological conditions, including dengue virus infection progression, chronic hepatitis C, and multiple cancer types. This dual functionality enhances the value of HGF in comprehensive research programs.

Expression System Advantages

Human cell-expressed recombinant HGF offers superior characteristics for clinical applications, including animal component-free production, tag-free formats, and native folding and maturation, which are particularly important when results must translate to therapeutic development.

Yes, recombinant human HGF can be used as a standard for quantification or calibration in ELISA assays, provided it is well-characterized and compatible with your assay system. Recombinant HGF is commonly used as a standard in commercial ELISA kits designed to quantify human HGF in biological samples.

Key considerations and supporting details:

• Assay Compatibility: Most quantitative sandwich ELISA kits for human HGF use recombinant human HGF as the standard to generate the calibration curve. These kits are validated to ensure that recombinant HGF produces a standard curve that is parallel to curves generated with natural HGF, allowing accurate quantification of both recombinant and endogenous HGF in samples.

• Standard Preparation: The recombinant HGF standard should be prepared according to the ELISA kit instructions, typically by serial dilution in the recommended diluent to cover the assay’s dynamic range. The concentration of HGF in unknown samples is then interpolated from the standard curve generated using the recombinant standard.

• Validation: It is important to confirm that your recombinant HGF is of high purity, correctly folded, and biologically active, as these factors affect its performance as a standard. Commercial ELISA kits often use NS0-expressed or similarly validated recombinant HGF for calibration.

• Matrix Effects: When quantifying HGF in complex biological matrices (e.g., serum, plasma), ensure that the recombinant standard behaves similarly to endogenous HGF in your assay system. Some kits recommend running internal controls or matrix-matched standards to account for potential matrix effects.

• Documentation: Always refer to the specific ELISA kit protocol for details on standard preparation and calibration procedures, as requirements may vary between kits.

Summary Table: Use of Recombinant Human HGF as ELISA Standard

In conclusion, recombinant human HGF is suitable and widely used as a standard for ELISA quantification, provided it is properly validated and compatible with your assay system.

Recombinant human HGF has been validated for a diverse range of applications across multiple research domains, as demonstrated in published literature.

Cell-Based Applications

Proliferation and Migration Studies

Recombinant HGF has been extensively validated for inducing cell proliferation and migration across various cell types. The protein demonstrates dose-dependent proliferation effects, with reported ED₅₀ values ranging from 0.5–3.5 ng/mL in certain epithelial cell lines. Additionally, HGF stimulates IL-11 secretion in human osteosarcoma cells with an ED₅₀ of less than 4.00 ng/mL.

Hepatocyte Culture and Expansion

A primary application involves in vitro culture and expansion of hepatocytes. HGF functions as a potent mitogen for mature hepatocytes in primary culture, making it valuable for hepatic tissue engineering and regenerative medicine approaches.

Tissue Engineering and Regeneration

Recombinant HGF has been validated for tissue development, regeneration, and organogenesis applications. Research has demonstrated its utility in biofabrication of synthetic human liver tissue with advanced programmable functions, as well as in promoting angiogenesis and supporting hematopoietic progenitor colony formation.

Therapeutic and Disease Model Applications

Liver Disease Models

Published research has validated recombinant HGF in mitigating various liver pathologies, including alcohol-induced liver injury, hepatic ischemia-reperfusion injury, and non-alcoholic steatohepatitis (NASH). Studies show that HGF treatment suppresses fibrogenic gene expression, reduces lipid accumulation, and decreases inflammatory markers in disease models.

Hair Regeneration

Recombinant HGF vectors have been validated for promoting hair follicle growth and regeneration through transfection of dermal fibroblasts, demonstrating improved follicle development and increased HGF receptor expression in animal models.

Analytical and Immunological Applications

Recombinant HGF has been validated for use in functional assays, ELISA, Western blotting, blocking assays, and immunohistochemistry. It also serves as an immunoassay standard and control reagent in various experimental protocols.

Clinical Translation

Early clinical investigations have employed recombinant human HGF protein for treating chronic leg ulcers, with ongoing phase-I and phase-I/II clinical trials examining intrathecal administration for amyotrophic lateral sclerosis and spinal cord injury.

To reconstitute and prepare Recombinant Human HGF (Hepatocyte Growth Factor) protein for cell culture experiments, follow these general guidelines based on manufacturer recommendations and best practices:

Reconstitution

1. Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
2. Reconstitute the protein in sterile, endotoxin-free PBS (pH 7.4). Most protocols recommend a concentration of 100 µg/mL (0.1 mg/mL) for initial reconstitution. Some suppliers suggest up to 0.2 mg/mL.
3. Add carrier protein (e.g., 0.1% endotoxin-free recombinant human serum albumin or BSA) to stabilize the HGF solution, especially for long-term storage or sensitive cell culture applications.
4. Gently mix by swirling or tapping the vial; avoid vigorous shaking to prevent denaturation.

Example Protocol

• For a vial containing 10 µg of HGF, add 100 µL of sterile PBS (with or without 0.1% HSA/BSA) to achieve a 100 µg/mL stock solution.
• For 20 µg, add 200 µL, etc.

Storage and Handling

• Short-term storage: Store reconstituted HGF at 2–8°C for up to 1 week if used soon.
• Long-term storage: Aliquot and store at –20°C to –70°C for up to 3 months. Avoid repeated freeze-thaw cycles.
• Unreconstituted (lyophilized): Store at –20°C to –70°C until use.

Preparation for Cell Culture

• Dilute the reconstituted HGF stock to the desired working concentration in your cell culture medium (typical working concentrations range from 10–100 ng/mL, depending on the cell type and application).
• Filter-sterilize the diluted solution if necessary (using a 0.22 µm filter).
• Add directly to the culture medium.

Additional Notes

• Always refer to the Certificate of Analysis (COA) or product datasheet for specific instructions, as formulations and recommendations may vary slightly between suppliers.
• Use endotoxin-free reagents and sterile technique to prevent contamination and ensure reproducibility in cell culture experiments.

This protocol ensures optimal activity and stability of Recombinant Human HGF for use in cell culture applications.