Cultrex BME
Cultrex Basement Membrane Extract (BME) is the trade name for a extracellular protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells and manufactured into a hydrogel by R&D Systems, a brand of Bio-Techne. Similar to Matrigel, this hydrogel is a natural extracellular matrix that mimics the complex extracellular environment within complex tissues. It is used as a general cell culture substrate across a wide variety of research applications.[1][2]
Composition
[edit]The major components of Cultrex BME include laminin, collagen, entactin/nidogen, and heparin sulfate proteoglycans. These components are found in the basement membrane extracellular matrix that forms an interface between stromal tissue and adjacent endothelial, epithelial, muscle, or neuronal cells. Also present in Cultrex BME are growth factors, however these can be reduced during production of the hydrogel. Reduced growth factor (RGF) Cultrex BME is commonly used for the culture of pluripotent stem cells and organoids.[citation needed]
Cell culture
[edit]Cultrex BME is gelatinous at 4 °C. The matrix proteins polymerize and solidify at temperatures above 18 °C. Ice-cold basement membrane extract can be dispensed directly onto plastic cell culture labware or it can be diluted in ice-cold phosphate buffered saline or cell culture media prior to dispensing.
Due to its heterogenous extracellular matrix protein composition, cells cultured using basement membrane extract show complex cellular behaviors that are difficult to reproduce under laboratory conditions. These include cell adhesion, migration, invasion, proliferation, differentiation, and endothelial cell tube formation.[3] EHS tumor-derived matrices, such as Cultrex BME, are used for a variety of cell culture applications, including angiogenesis,[4] spheroid formation, organoid culture, pluripotent stem cell culture, xenograft, and in vivo and in vitro tumor modeling[5][6]
Pluripotent stem cells
[edit]Cultrex BME is used as an attachment substrate to promote the expansion and maintenance of induced pluripotent stem cells and embryonic stem cells in the absence of feeder cells.[7] The extracellular matrix proteins that compose the basement membrane extract are needed to maintain stem cells in an undifferentiated state during prolonged cell culture. For expanding stem cells, the hydrogel is commonly diluted prior to coating cell culture plasticware. Concentrated Cultrex BME is often used during cell differentiation or transitioning pluripotent stem cells into organoid or spheroid culture.
Organoid and 3D cell culture
[edit]Proliferation and differentiation of spheroids and organoids, derived from induced pluripotent stem cells or adult tissue-specific stem cells, is supported by EHS-derived hydrogels such as Cultrex BME. EHS-derived hydrogels have been used to generate a variety of organoids, including intestinal, lung, liver, and snake venom gland[8][9][10]
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References
[edit]- ^ Benton, G.; George, J.; Kleinman, H. K.; Arnaoutova, I. P. (October 2009). "Advancing science and technology via 3D culture on basement membrane matrix". Journal of Cellular Physiology. 221 (1): 18–25. doi:10.1002/jcp.21832. ISSN 1097-4652. PMID 19492404. S2CID 37221327.
- ^ Kleinman, Hynda K.; Martin, George R. (October 2005). "Matrigel: basement membrane matrix with biological activity". Seminars in Cancer Biology. 15 (5): 378–386. doi:10.1016/j.semcancer.2005.05.004. ISSN 1044-579X. PMID 15975825.
- ^ Arnaoutova, Irina; George, Jay; Kleinman, Hynda K.; Benton, Gabriel (2009). "The endothelial cell tube formation assay on basement membrane turns 20: state of the science and the art". Angiogenesis. 12 (3): 267–274. doi:10.1007/s10456-009-9146-4. ISSN 1573-7209. PMID 19399631. S2CID 23510613.
- ^ Arnaoutova, Irina; Kleinman, Hynda K. (April 2010). "In vitro angiogenesis: endothelial cell tube formation on gelled basement membrane extract". Nature Protocols. 5 (4): 628–635. doi:10.1038/nprot.2010.6. ISSN 1750-2799. PMID 20224563. S2CID 7720444.
- ^ Benton, Gabriel; Kleinman, Hynda K.; George, Jay; Arnaoutova, Irina (2011-04-15). "Multiple uses of basement membrane-like matrix (BME/Matrigel) in vitro and in vivo with cancer cells". International Journal of Cancer. 128 (8): 1751–1757. doi:10.1002/ijc.25781. ISSN 1097-0215. PMID 21344372. S2CID 24269013.
- ^ Benton, Gabriel; Arnaoutova, Irina; George, Jay; Kleinman, Hynda K.; Koblinski, Jennifer (2014-12-15). "Matrigel: from discovery and ECM mimicry to assays and models for cancer research". Advanced Drug Delivery Reviews. 79–80: 3–18. doi:10.1016/j.addr.2014.06.005. ISSN 1872-8294. PMID 24997339.
- ^ Arnaoutova, Irina; George, Jay; Kleinman, Hynda K.; Benton, Gabriel (March 2012). "Basement membrane matrix (BME) has multiple uses with stem cells". Stem Cell Reviews and Reports. 8 (1): 163–169. doi:10.1007/s12015-011-9278-y. ISSN 2629-3277. PMID 21655946. S2CID 2913671.
- ^ Co, Julia Y.; Margalef-Català, Mar; Li, Xingnan; Mah, Amanda T.; Kuo, Calvin J.; Monack, Denise M.; Amieva, Manuel R. (26 February 2019). "Controlling Epithelial Polarity: A Human Enteroid Model for Host-Pathogen Interactions". Cell Reports. 26 (9): 2509–2520.e4. doi:10.1016/j.celrep.2019.01.108. ISSN 2211-1247. PMC 6391775. PMID 30811997.
- ^ Sachs, Norman; Papaspyropoulos, Angelos; Zomer-van Ommen, Domenique D.; Heo, Inha; Böttinger, Lena; Klay, Dymph; Weeber, Fleur; Huelsz-Prince, Guizela; Iakobachvili, Nino; Amatngalim, Gimano D.; de Ligt, Joep (15 February 2019). "Long-term expanding human airway organoids for disease modeling". The EMBO Journal. 38 (4). doi:10.15252/embj.2018100300. ISSN 1460-2075. PMC 6376275. PMID 30643021.
- ^ Post, Yorick; Puschhof, Jens; Beumer, Joep; Kerkkamp, Harald M.; de Bakker, Merijn A. G.; Slagboom, Julien; de Barbanson, Buys; Wevers, Nienke R.; Spijkers, Xandor M.; Olivier, Thomas; Kazandjian, Taline D. (2020-01-23). "Snake Venom Gland Organoids". Cell. 180 (2): 233–247.e21. doi:10.1016/j.cell.2019.11.038. hdl:1871.1/ff5336a2-3e02-4d9c-8997-2e4396580d89. ISSN 1097-4172. PMID 31978343. S2CID 210860537.