Discovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)
/lifesciences
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GUIDELINES FOR USE
PRODUCT: Corning ® Matrigel ® Bament Membrane Matrix High Concentration, 10 ml vial CATALOG NUMBER: 354248
BACKGROUND: Bament membranes are thin extracellular matrices underlying cells in vivo . Corning
Matrigel Matrix High Concentration (HC) is a solubilized bament membrane
preparation extracted from the Engelbreth-Holm-Swarm (EHS) mou sarcoma, a
缓存怎么清理
tumor rich in extracellular matrix proteins. Its major component is laminin, followed
拍照的英文
by collagen IV, heparan sulfate proteoglycans, entactin/nidogen.1,2 Corning Matrigel
Matrix HC also contains TGF-beta, epidermal growth factor, insulin-like growth
长征的历史意义factor, fibroblast growth factor, tissue plasminogen activator,3,4 and other growth
factors which occur naturally in the EHS tumor. Corning Matrigel Matrix HC is
effective for the attachment and differentiation of both normal and transformed
anchorage dependent epithelioid and other cell types. The include neurons,5,6
hepatocytes,7 Sertoli cells,8,9 chick lens,10 and vascular endothelial cells.11 Corning
Matrigel Matrix HC will influence gene expression in adult rat hepatocytes 12,13 as well
as three dimensional culture in mou 14-17 and human 18,19 mammary epithelial cells. It
is the basis for veral types of tumor cell invasion assays,20,21 will support in vivo
peripheral nerve regeneration,22-24 and provides the substrate necessary for the study
of angiogenesis both in vitro 25,26 and in vivo .27-29 Corning Matrigel Matrix HC also
supports in vivo propagation of human tumors in immunosupresd mice.30-32 For
further information, go to our website /lifesciences.
SOURCE:
Engelbreth-Holm-Swarm (EHS) Mou Tumor FORMULATION:
保险制度
Dulbecco's Modified Eagle's Medium with 50 g/ml gentamycin
Corning Matrigel Matrix HC is compatible with all culture media STABILITY:
Stable for a minimum of three months from day of shipment when stored at -20 C
KEEP FROZEN
RECONSTITUTION
AND USE: Color variations may occur in frozen or thawed vials of Corning Matrigel Matrix HC,
ranging from straw yellow to dark red due to the interaction of carbon dioxide with the
bicarbonate buffer and phenol red. Variation in color is normal, does not affect product
efficacy, and will disappear upon equilibration with 5% CO 2.
Once Corning Matrigel Matrix HC is thawed, swirl vial to be sure that material is
evenly disperd. Handle using sterile technique. Place thawed vial of Corning Matrigel
Matrix HC in sterile area, spray top of vial with 70% ETOH and air dry. Corning
Matrigel Matrix HC may be gently pipetted using a pre-cooled pipette to ensure
homogeneity.
Corning Matrigel Matrix HC may be ud as a thin gel layer (0.5mm), with cells
plated on top. Cells may also be cultured inside the Corning Matrigel Matrix HC,
using a 1 mm layer. Extensive dilution will result in a thin, non-gelled protein layer.
This may be uful for cell attachment, but may not be as effective in differentiation
studies. Corning Matrigel Matrix HC can be ud to asss in vivo angiogenic activity
of different compounds by subcutaneous injection into mice (Corning Matrigel Plug
Assay).2,8,25 The high protein
李连杰身价Discovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)
/lifesciences羽毛球竞赛规则
For Rearch U Only. Not for u in diagnostic or therapeutic procedures.
concentration augments the growth of tumors and also allows the Corning ® Matrigel ®
Plug to maintain its integrity after injection. This keeps the injected tumor and/or angiogenic compounds localized for in situ analysis and/or future excision.
Dispen remaining material into appropriate aliquots, using pre-cooled tubes, and
refreeze immediately. Avoid multiple freeze thaws. DO NOT STORE IN FROST-
FREE FREEZER.
CAUTION :
Corning Matrigel Matrix HC will gel rapidly at 22o C to 35o C. Thaw overnight at 4o C on ice (Matrigel may gel at slightly elevated temperatures in a refrigerator). Keep product on ice before u, and u pre-cooled pipettes, tips, and tubes when preparing Corning Matrigel Matrix HC for u. Gelled Corning Matrigel Matrix HC may be re-liquified if placed at 4°C on ice for 24-48 hours.
INJECTION PROTOCOL:
1. It is critical to keep the Corning Matrigel Matrix HC and the Corning Matrigel/Cell suspension as cold as
possible, without freezing, prior to injecting into the mice. It is very important to keep the Corning Matrigel and the Corning Matrigel/Cell suspension as asceptic as possible throughout the procedure.
2. For each recipient mou, mix cells (2 x 105 or greater) and Corning Matrigel Matrix HC together in a final
volume of 0.5 ml on ice.
3. The cells should be in as small a volume as possible. Typically, 250 l ice cold medium containing 2 x 106
cells/ml is mixed with 250 l ice cold Corning Matrigel Matrix HC.
4. Inject the cells subcutaneously in athymic mice using a 19G needle for tissue samples and a 23G needle for
cultured cells. The injections should be done quickly to prevent the Matrigel from solidifying.
5. Rotate the syringe when withdrawing to prevent leakage. The needles will need to be changed frequently due
to blockage.
NOTE: For more details on this application go /lifesciences to access CLS-DL-CC-036 (Technical Bulletin 455: Methods for Implantation of Corning Matrigel Matrix into Mice and Tissue Fixation).
CELL RECOVERY:
Dispa (Catalog No. 354235), Corning Cell Recovery Solution (Catalog No. 354253)
Most efficient recovery of cells growing on Corning Matrigel Matrix HC is accomplished using Corning Cell
Recovery Solution that depolymerizes the Matrigel Matrix within 7 hours on ice or with Dispa, a metalloenzyme which gently releas the cells allowing for continuous culture.
REFERENCES:
1.
Kleinman, H.K., et al., Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma, Biochemistry, 21:6188 (1982).
2. Kleinman, H.K., et al., Bament membrane complexes with biological activity, Biochemistry, 25:312 (1986).
3. Vukicevic, S., et al., Identification of multiple active growth factors in bament membrane Matrigel suggests caution in interpretation of
cellular activity related to extracellular activity related to extracellular matrix components, Experimental Cell Rearch, 202:1 (1992).
4. McGuire, P.G. and Seeds, N.W., The interaction of plasminogen activator with a reconstituted bament membrane matrix and extracellular
macromolecules produced by cultured epithelial cells, J. Cell. Biochem., 40:215 (1989).
Discovery Labware, Inc ., Two Oak Park, Bedford, MA 01730, Tel: 1.978.442.2200 (U.S.)
/lifesciences
For Rearch U Only. Not for u in diagnostic or therapeutic procedures.
5.
经典散文欣赏50篇
Biederer, T. and Scheiffele, P., Mixed-culture assays for analyzing neuronal synap formation, Nature Protocols, 2(3):670 (2007). 6. Li, Y., et al., Esntial Role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor, Nature, 434:894
(2005).
7. Bi, Y., et al., U of cryoprerved human hepatocytes in sandwich culture to measure hepatobiliary transport, Drug Metabo. and Dispos.,
34(9):1658 (2006).
8. Hadley, M.A., et al., Extracellular matrix regulates rtoli cell differentiation, testicular cord formation, and germ cell development in vitro, J.
Cell Biol., 101:1511 (1985).
9. Yu, X., et al., Esntial role of extracellular matrix (ECM) overlay in establishing the functional integrity of primary neonatal rat rtoli
cell/gonocyte co-cultures: An improved in vitro model for asssment of male reproductive toxicity, Toxilogical Sciences, 84(2):378 (2005).
10. Ireland, M.E., Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens, 16(8):838 (1997).
11. McGuire, P.G., and Orkin, R.W., A simple procedure to culture and passage endothelial cells from large vesls of small animals,
Biotechniques, 5(6):456 (1987).
12. Bisl, D.M., et al., Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in
normal rat liver, J. Clinical Invest., 79:801 (1987).
13. Page, J.L., et al., Gene expression profiling of extracellular matrix as an effector of human hepatocyte phenotype in primary cell culture,
Toxilogical Sciences, 97(2):384 (2007).
14 Li, M.L., et al., Influence of a reconstituted bament membrane and its components on cain gene expression and cretion in mou
mammary epithelial cells, Proc. Nat. Acad. Sci. USA, 84:136 (1987).
15 Barcellof, M.H., et al., Functional differentiation and aveolar morphogenesis of primary mammary cultures on reconstituted bament
membrane, Development, 105:223 (1989).
16. Roskelley, C.D., et al., Extracellular matrix-dependent tissue-specific gene expression in mammary epithelial cells requires both physical and
biochemical signal transduction, Proc. Nat. Acad. Sci. USA, 91(26):12378 (1994).
电磁炉加热原理17. Xu, R., et al., Extracellular matrix-regulated gene expression requires cooperation of SWI/SNF and transcription factors, J. Biol. Chem.,
282(20):14992 (2007).
18. Debnath, J., et al., Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional bament membrane
cultures, Methods, 30(3):256 (2003).
19. Muthuswamy, S.K., et al., ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini, Nat. Cell Biol.,
3(9):785 (2001).
20. Terranova, V.P., et al., U of a reconstituted bament membrane to measure cell invasiveness and lect for highly invasive tumor cells, Proc.
Nat. Acad. Sci. USA, 83:465 (1986).
21. Albini, A., et al., A rapid in vitro assay for quantitating the invasive potential of tumor cells, Cancer Rearch, 47:3239 (1987).
22. Madison, R., et al., Incread rate of peripheral nerve regeneration using bioresorbable nerve guides and laminin containing gel, Exp. Neurology, 88:767 (1985).
23. Xu, X.M., et al., Axonal regeneration into Schwann cell-eded guidance channels grafted into trancted adult rat spinal cord, J. Comp. Neurol., 351(1):145 (1994).
24. Fouad, K., et al., Combining schwann cell bridges and olfactory-ensheathing glia grafts with chondroitina promotes locomotor recovery after
complete tranction of the spinal cord, The Journal of Neuroscience, 25(5):1169 (2005).
25. Kubota, Y., et al., Role of laminin and bament membrane in the morphological differentiation of human endothelial cells into capillary-like
structures, J. Cell Biol., 107:1589 (1988).
26. Maeshima, Y., et al., Identification of the anti-angiogenic site within vascular bament membrane-derived Tumstatin, J. Biol. Chem.,
276(18):15240 (2001).
27. Passaniti, A., et al., A simple, quantitative method for asssing angiogenesis and anti-angiogenic agents using reconstituted bament
membrane, heparin, and fibroblast growth factor , Lab Invest., 67:519 (1992).
28. Isaji, M., et al., Tranilast inhibits the proliferation, chemotaxis and tube formation of human microvascular endothelial cells in vitro and
angiogenesis in vivo, British Journal of Pharmacology, 122:1061 (1997).
29. Kisucka, J., et al., Platelets and platelet adhesion support angiogenesis while preventing excessive hemorrhage, Proc. Nat. Acad. Sci. USA,
103(4):855 (2006).
30. Albini, A., et al., Matrigel promotes retinoblastoma cell growth in vitro and in vivo, Int. J. Cancer, 52(2):234 (1992).
31. Yue, W., et al., MCF-7 human breast carcinomas in nude mice as a model for evaluating aromata inhibitors, J. Steroid Biochem. Molec. Biol.,
44(4-6):671 (1993).
32. Angelucci, A., et al., Suppression of EGF-R signaling reduces the incidence of prostate cancer metastasis in nude mice, Endocrine-Related
Cancer, 13(1):197 (2006).
CALIFORNIA PROPOSITION 65 NOTICE
WARNING: This product contains a chemical known to the state of California to cau cancer.
Component: Chloroform
