Collagen I, bovine - FAQs

I型胶原是以20mM乙酸溶液的液体形式来提供的。如果在准备过程中遇到困难，可先按1:2的比例将胶原稀释于20mM，pH 3.5的乙酸中，之后再按所用的实验方案中的说明进一步稀释。

可用的酶包括：Gibco TrypLE  Express，Gibco TrypLE  Select或Gibco StemPro Accutase 细胞解离试剂（Cell Dissociation Reagent）。

以下细胞类型通常培养于I型胶原中：

- 内皮细胞（包括HUVEC）
- 成纤维细胞
- 肝细胞
- 软骨细胞
- 破骨细胞
- 成骨细胞
- 角质形成细胞
- 角膜上皮

其关键应用列举如下：

- I型胶原能够在体外诱导微血管内皮细胞形成纺锤状形态，并排列为类似实线的聚集体。在I型胶原上进行培养时，血管内皮细胞也能够形成类似血管的管状结构。因此，I型胶原也可用于体外的血管生成实验。
- 乳腺癌干细胞在I型胶原上培养时可发生分化。
- I型胶原同样已被用于原代结肠癌细胞系的培养；小鼠肝脏前体细胞和大鼠的胰岛细胞也可培养于3D的I型胶原中。
- I型胶原可在细胞侵润实验分析中作为屏障，也已被用于细胞粘附的相关研究。

Gibco Geltrex 基质与胶原（大鼠尾与牛）可作为包被溶液或3D凝胶基质来使用。CELLstart 底物作为一种不含异外源性物质的包被基质，专为ESC的相关应用而研发（作为动物源性的Gibco Geltrex 基质或动物源性的Matrigel 基质的一种替代品底物）。AlgiMatrix 基质是为一种3D支架类的基质，它不支持并不为细胞提供粘附的基础，但而是为细胞球生长提供一个良好的生长环境，后续应用中也可方便易于细胞球的收取操作用于后续应用。

The polystyrene that culture flasks are made of tends to have a negative charge. Many cell types will still attach because the plasticware is treated in a way to make them hydrophilic. However cell types that do not attach well to the plastic, or are grown in medium with little or no serum protein, will attach better if the plastic is coated with a protein/polymer which has an overall positive charge.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

An alternative for PureCol, Type I Collagen Solution, 3 mg/ml (Bovine) (Cat. No. 5005) would be Collagen I, bovine (Cat. No. A1064401), which can be found here:

Collagen I, bovine (Cat. No. A1064401)

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Unfortunately, we do not have an animal-origin-free alternative for the Coating Matrix Kit Protein (Cat. No. R011K).However, we offer Rat or Bovine collagen that can be used as coating matrices. Please see the product pages linked below:

Collagen I, bovine (Cat. No. A1064401)

Collagen I, rat tail (Cat. No. A1048301)

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Follow the gelling procedure below to change Collagen I, bovine (Cat. No. A1064401) from liquid to gel.

Gelling procedures (perform under sterile conditions):

1. Place the following on ice:

    Collagen (5 mg/mL)
    Phosphate buffer saline (10X) or Medium 199 (10X)
    Sterile distilled water
    Sterile 1N NaOH

2. Determine the concentration and final volume of collagen needed for experimentation. For optimal gel formation, a concentration of 3 or 4 mg/mL is recommended.

3. Determine the amount of reagents needed to achieve the desired collagen concentration in 1X PBS or Medium 199 with normal osmolality and neutral pH.

     - V = total volume of collagen gel desired

     - Volume of collagen needed (V1) = (Final conc. of collagen) x ( total volume (V))/Initial concentration of collagen

     - Volume of 10X PBS needed (V2) = Total volume (V)/10

     - Volume of 1N NaOH needed (V3) = (total volume of collagen needed (V1)) x 0.025

     - Volume of dH2O needed (V4) = Total volume – (calculated volumes from steps V1 + V2 + V3)

4. Mix dH2O, 1N NaOH, and 10X PBS in a sterile tube.

5. Rinse the gel with 1X PBS or cell culture medium before seeding cells.

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An alternative for Collagen I, rat tail (Cat. No A1048301) is Collagen I, bovine (Cat. No A1064401). While the source of Collagen I is different, they can both be prepared as a clear gel providing a 3D matrix or surface coating on tissue culture plates as a substrate for culturing primary cells such as keratinocytes and hepatocytes.

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Collagen I, bovine (Cat. No. A1064401) is shipped in 20 mM acetic acid.

Collagen I is provided as a liquid, suspended in 20 mM acetic acid solution. If difficulties are encountered with the preparation, the collagen can be diluted in 1:2, first in 20 mM acetic acid, pH 3.5 before further diluting according to the protocol being used.

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Enzymes that can be used include: Gibco TrypLE Express Enzyme, Gibco TrypLE Select Enzyme, or Gibco StemPro Accutase Cell Dissociation Reagent.

Find additional tips, troubleshooting help, and resources within our Extracellular Matrices (ECM) Support Center.

The following cell types are commonly cultured in collagen I:

- Endothelial cells (including HUVEC)
- Fibroblasts
- Hepatocytes
- Chondrocytes
- Osteoclasts
- Osteoblasts
- Keratinocytes
- Corneal epithelium

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The key applications include the following:

- Collagen I induces microvascular endothelial cells to adopt a spindle-shaped morphology, in vitro, and to align into solid cord-like assemblies. Vascular endothelial cells can also form vessel-like, tubular structures when cultured on collagen I. Collagen I can be used for in vitro angiogenesis assays.
- Breast cancer stem cells can undergo differentiation when cultured on collagen I.
- Collagen I has been used for the culture of primary colon carcinoma cell lines; mouse liver progenitor cells have been cultured in 3D collagen I and rat pancreatic islets.
- Collagen I can act as a barrier in cell invasion assays, and has been used to study cell adhesion.

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Gibco Geltrex Matrix and collagen (rat tail and bovine) can be used as either a coating solution or a 3D gel matrix. CELLstart Substrate was developed to be used as a xeno-free coating matrix for only ESC applications (as a substitute for Gibco Geltrex Matrix or Matrigel Matrix, which are of animal origin). AlgiMatrix Matrix is a 3D scaffold-type matrix that does not support cell attachment but does provide a good environment for growing spheroids that can be easily harvested for downstream applications.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.