Product Number 3446-005-01
3D Culture is a cell culture method that provides cells with the necessary structure and signaling cues to direct reconstruction of the tissue architecture, and these methods provide physiologically predictive in vitro models for evaluating development and disease. Normal cells assemble into organoids that structurally resemble their emanating tissues, exhibit a polarized morphology, undergo cell cycle regulation, and produce tissue specific proteins1-6. While cancer cells assemble into tumor-like structures, lacking an organized architecture or cell cycle regulation and exhibiting tumor-specific markers depending on the extend of malignancy7-9. To aid in the advancement of this technology, we present Cultrex® 3-D Culture Matrix™ RGF BME which is the first Basement Membrane Extract produced and qualified specifically for use in 3D culture studies. The 3-D Culture Matrix™ RGF BME provides the foundation for cells to grow in three dimensions allowing for the formation of structures in vitro. To provide the most standardized basement membrane extract for use in 3D cultures, a special process is employed to reduce growth factors and manufacture matrix at a standard concentration of approximately 15 mg/mL. This product is then evaluated in a 3D culture to validate efficacy.
Concentration: Please refer to lot specific data
Purity: ≥90% (SDS-PAGE)
Source: Murine Engelbreth-Holm-Swarm (EHS) tumor
Storage buffer: Dulbecco’s Modified Eagle’s medium without phenol red (Product No. D5030), with 10 μg/mL gentamicin sulfate (Product No. G1264)
For Research Use Only. Not for use in diagnostic procedures.
The physical, chemical, and toxicological properties of these products may not yet have been fully investigated; therefore, we recommend the use of gloves, lab coats, and eye protection while using these chemical reagents.
1. Functional Assays
Cell Attachment – Tested for the ability to promote cell attachment and spreading of MG63 human osteosarcoma cells.
3D culture – Laminin I promotes attachment and growth of a human epithelial cell line derived from mammary gland (MCF-10A) and human prostate (PC-3), and in the presence of assay medium, these cell lines differentiate into acinar structures.
2. Sterility Testing
No bacterial or fungal growth detected after incubation at 37 ⁰C for 14 days following USP XXIV Chapter 71 sterility test
No mycoplasma contamination detected by PCR
Endotoxin concentration ≤20 EU/mL by LAL assay
1. Laminin I forms a firm gel at neutral pH and 37 ⁰C at 6 mg/mL.
Store at – 20 ⁰C in a manual defrost freezer or at – 80 ⁰C.
This procedure must be conducted in an aseptic environment, such as a laminar flow hood or clean room, using aseptic technique to prevent contamination.
Recommendations for analysis
3-D Culture of MCF-10A Breast Cancer Cells using 3-D Culture Matrix™ Laminin I for 15 days in the presence of 2% Liminin I in Assay Medium, and stained using A) CPA Dye 2, and B) SYBR® Green (nuclear) and propidium iodine (dead or necrotic).
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