Cultrex 3-D Culture Matrix Laminin I
Cultrex 3-D Culture Matrix Laminin I Summary
Why Use Cultrex 3-D Culture Matrix Laminin I?
Cultrex 3-D Culture Matrix Laminin I is purified from Engelbreth-Holm-Swarm (EHS) tumor and is provided at a high concentration that is capable of polymerizing to form a hydrogel at 37°C. Laminin I is a major component of the basement membrane which is a continuous sheet of specialized extracellular matrix that forms an interface between endothelial, epithelial, muscle, or neuronal cells and their adjacent stroma and that plays an essential role in tissue organization by influencing cell adhesion, migration, proliferation, and differentiation. Cultrex 3-D Culture Matrix Laminin I is a purified basement membrane protein that has been developed, produced and qualified specifically for use in 3-D culture studies. Cultrex 3-D Culture Matrix Laminin I may also be used as to supplement customized hydrogel or medium formulations for cell culture.
Cell Attachment - Tested for the ability to Supports cell attachment and spreading of MG63 human osteosarcoma cells.
For research use only. Not for diagnositic use.
Citations for Cultrex 3-D Culture Matrix Laminin I
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Ythdf2-mediated m6A mRNA clearance modulates neural development in mice
Authors: M Li, X Zhao, W Wang, H Shi, Q Pan, Z Lu, SP Perez, R Suganthan, C He, M Bjørås, A Klungland
Genome Biol., 2018;19(1):69. 2018
Coordination of epithelial branching and salivary gland lumen formation by Wnt and FGF signals.
Authors: Patel N, Sharpe PT, Miletich I
Dev. Biol., 2011;358(1):156-67. 2011
What type of analysis is typically applied for organoid or 3-D cell cultures?
Within the organoid, spheroid, or 3-D culture, cells may be assessed for morphology, apical/basal polarity, protein localization, and relative proliferation. In addition, cells may be isolated from the 3-D culture and evaluated for levels of RNA and protein expression, as well as modifications to DNA.
What is the recommended working concentration for Cultrex Laminin I?
The recommended working concentration for thin coating is 0.05-10 µg/cm2 . However, conditions must be optimized for each cell line or model. Cultrex Laminin I should be used at 6 mg/mL for 3-D culture applications.
What is Laminin I?
Laminin I is a major component of extracellular matrix. Cultrex Laminin I is purified from murine EHS sarcoma. It is composed of α1β1γ1 chains with a total MW of 800,000 Da. Cultrex Laminin I increases cell adhesion, migration, growth, differentiation, neurite outgrowth, protease production, and malignancy. The response is dependent on cell type.
What are 3-D cultures?
3-D cultures are in vitro cultures where immortalized cell lines, primary cell lines, stem cells, or tissue explants are placed within hydrogel matrices, such as Cultrex Basement Membrane Extract, that mimic in vivo cell environments and allow cells to proliferate in three dimensions.
What is the advantage of 3-D culture over traditional 2-D culture?
While 2-D culture has been used for studying many aspects of cell function and behavior, the tissue-culture treated plastic environment is unlike anything found within living organisms. As a result, cells in 2-D culture exhibit altered morphology, function, proliferation, and gene expression when compared to their emanating tissues. By placing these cells in a 3-D environment, they assume biological and biochemical characteristics similar to what is observed in vivo.
What are the variables associated with 3-D culture?
The major variables associated with 3-D culture are cell type, cell seeding density, composition of hydrogel, thickness of hydrogel, stiffness of hydrogel, composition of cell culture medium, and time of culture.
What are the different types of 3-D culture?
The two principal methods for performing 3-D culture are the top assay and embedded assay. For the top assay, cells are seeded on a thick gel of Cultrex Basement Membrane Extract (BME) or Extracellular Matrix Protein. A thin overlay of cell culture medium is then applied to the cells. For the embedded assay, cells are resuspended within a thick gel of Cultrex BME or ECM and the culture media is applied on top. The top assay is easier to setup, to control seeding densities, and to keep cells within one focal plane for analysis.
Which matrix should I use for 3-D culture?
Choice of matrix should correspond to the environment that you wish to recapitulate. Cultrex Basement Membrane Extract (BME) will recapitulate the basal lamina, which underlie most cells of epithelial or endothelial origin. Cultrex Collagen I is the major constituent of connective tissue, and is commonly inhabited by stationary cells, such as fibrocytes and adipose cells, as well as migrating cells, such as mast cells, macrophages, monocytes, lymphocytes, plasma cells, and eosinophils.
How should cells be cultured prior to setting up the 3-D culture?
Cells need to be healthy and actively dividing in 2-D culture. Cells should be passaged two or three times after resuspension from cryopreservation, and they should never surpass 80% confluency during each passage. Cells should also be assessed for viability using trypan blue, and they should exhibit less than 5% staining.
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