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 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.

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.

Cultrex 3-D Culture Matrix Laminin I Best Seller

Name: Cultrex 3-D Culture Matrix Laminin I
Brand: R&D Systems
SKU: 3446-005-01

R&D Systems, part of Bio-Techne | Catalog # 3446-005-01

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Key Product Details

Features:

Cultrex 3-D Culture Matrix Laminin I is an extracellular matrix hydrogel that directs cells to grow in three dimensions and assemble into organotypic structures in vitro.

Technical Data
Citations / Reviews / FAQs

Summary for Cultrex 3-D Culture Matrix Laminin I

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.

Negative for the presence of bacteria and fungi.

Product Specifications for Cultrex 3-D Culture Matrix Laminin I

Species	Mouse
Source	Murine Engelbreth-Holm-Swarm (EHS) tumor
Protein Concentration	6 mg/mL
Endotoxin level	≤ 20 EU/mL by Limulus Amoebocyte Lysate (LAL) assay
Sterility Testing	No bacterial or fungal growth detected following 14 days in culture
Cell Culture Testing	3-D Culture - Laminin I Supports differentiation of a human epithelial cell line derived from mammary gland (MCF-10A) into acinar structures. Cell Attachment - Tested for the ability to Supports cell attachment and spreading of MG63 human osteosarcoma cells.

Preparation & Storage

Stability	Product is stable for a minimum of 3 months from date of shipment. See lot specific Certificate of Analysis for expiration date.
Shipping Conditions	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended on the product label.
Storage	Store at ≤ -20 °C in a manual defrost freezer. For optimal stability, store at ≤ -70 °C. Avoid freeze-thaw cycles

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Cultrex 3-D Culture Matrix Laminin I Best Seller

R&D Systems, part of Bio-Techne | Catalog # 3446-005-01

Key Product Details

Summary for Cultrex 3-D Culture Matrix Laminin I

Why Use Cultrex 3-D Culture Matrix Laminin I?

Product Specifications for Cultrex 3-D Culture Matrix Laminin I

Preparation & Storage

Customer Reviews for Cultrex 3-D Culture Matrix Laminin I

FAQs for Cultrex 3-D Culture Matrix Laminin I

Product Documents for Cultrex 3-D Culture Matrix Laminin I

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