Collagen-based Cell Contraction Assay Kit，IC-6580
Product Introduction
The Collagen-based Cell Contraction Assay provides a three-dimensional (3D) culture system to quantitatively measure cell-mediated collagen gel contraction. This in vitro model mimics tissue remodeling and wound healing processes, allowing researchers to evaluate the contractile capacity of various cell types, including fibroblasts, smooth muscle cells, and epithelial cells.
Cells embedded within a hydrated type I collagen lattice reorganize the matrix, reducing gel diameter and volume over time. Contraction is typically quantified by measuring gel area or weight, enabling assessment of:
- Drug/compound effects on contractility
- Mechanotransduction pathways (e.g., Rho/ROCK, MLCK)
- Cell-matrix interactions
Product Features
Physiologically relevant：3D matrix mimics native extracellular environment.
High reproducibility：Standardized collagen concentration and neutralization buffer.
Flexible format：Suitable for 24- or 48-well plates; easy scaling.
Quantitative：Measure gel diameter, area, or weight; optional time-lapse imaging.
Compatible：Works with primary cells, cell lines, and co-culture systems.
Kit Components (Example –check your lot)
- Type I Collagen Solution (e.g., 3 mg/mL in 0.1% acetic acid)
- 10× PBS or Neutralization Buffer
- 0.1 N NaOH (if required)
- Cell culture medium (e.g., DMEM, serum-free or with FBS – user supplied)
- 24-well culture plates (non-adherent or pre-coated recommended)
Assay Protocol (For Reference Only)
Note: Optimize cell number and collagen concentration for your specific cell type.
A. Preparation (Day 0)
1. Keep collagen solution on ice to prevent premature gelling.
2. Prepare collagen mix (per 1 mL gel, for triplicate wells of a 24-well plate – adjust as needed):
|
Component |
Volume (μL) |
|
Collagen solution (3 mg/mL) |
500 |
|
5× or 10× Medium/Neutralization buffer |
200 (for 5×) |
|
0.1 N NaOH (to adjust pH to ~7.4) |
20–30 (dropwise) |
|
Cell suspension (in culture medium) |
Variable (e.g., 200 μL) |
|
Sterile H₂O or 1× medium |
To final 1000 μL |
Final collagen concentration typically 1–2 mg/mL.
Final cell density: e.g., 1–5 × 10⁵ cells/mL for fibroblasts.
3. Mix gently – avoid air bubbles.
B. Gel Casting
4. Add 500 μL of collagen-cell mixture per well in a 24-well plate.
5. Incubate at 37℃, 5% CO₂ for 30–60 min until gel solidifies.
6. Overlay with 500 μL pre-warmed culture medium (with or without test compounds).
C. Contraction Measurement
7. Detach gels from well walls using a sterile pipette tip or needle along the perimeter.
8. Incubate for 24–72 hours (depending on cell contraction speed).
9. Quantify contraction:
- Area method: Photograph gels and measure diameter/area using ImageJ.
- Weight method: Carefully transfer gel to pre-weighed tube; measure wet weight.
- Calculate Contraction Index = (Initial area – Final area) / Initial area × 100%.
Important Precautions
- Avoid gel detachment during medium changes – add medium slowly against the wall.
- Do not vortex collagen solution – this causes fiber breakage and weak gels.
- pH is critical – too acidic or basic prevents gelling. Use phenol red as indicator (orange = neutral).
- Use low-adhesion plates or coat wells with BSA/agarose to prevent gel sticking.
- Control conditions must include:
- Cell-free collagen gel (negative contraction control)
- Positive control (e.g., 10% FBS or PDGF)
- Serum-free conditions may slow contraction; optimize accordingly.
Frequently Asked Questions (FAQ)
Q1: My collagen gel does not polymerize – why?
A: Most common causes:
- Incorrect pH (should be ~7.4).
- Temperature too low – gels must be at 37℃.
- Low collagen concentration (<0.5 mg/mL).
- Expired or denatured collagen.
Q2: Gels detach from the plate spontaneously before I release them.
A: Use non-adherent plates or pre-coat with 1% BSA. If using standard TC-treated plates, gently loosen gels at 30–60 min after polymerization.
Q3: How do I normalize contraction data between experiments?
A: Express contraction as percentage of initial gel area. Include an internal control (e.g., 10% FBS) in every experiment. Use triplicate wells per condition.
Q4: Can I use this assay for high-throughput screening?
A: Yes, adapt to 96-well plates with robotic liquid handling. Gel size is smaller – measure using automated image analysis or fluorescent bead displacement.
Q5: My cells contract too fast (<6 hours) – how to slow down?
A: Reduce cell density, increase collagen concentration (e.g., 2–3 mg/mL), or use serum-free medium with low mitogens.
Q6: Can I fix and stain gels after contraction?
A: Yes. Fix with 4% paraformaldehyde (30 min, RT). Stain F-actin (phalloidin) or nuclei (DAPI). For histology, paraffin-embed after fixation.
Product Statement
This product is for research use only. For storage of precious samples, please conduct small-scale pilot tests before large-scale experiments. Product after-sales service is limited to the product itself and does not include any other liability. Please be advised.
This product is for research use only. Not for use in clinical diagnosis or therapeutic procedures.
Order Information
|
Cat./REF. |
Size |
Price($） |
Price(�) |
Price(￥/CNY） |
Price(￥/JYP） |
|
IC-6580 |
24Assays |
$538.50 |
� 646.20 |
￥5,385.00 |
￥107,161.50 |
