Frequently Asked Questions
Is MatriMix the same as the mouse tumor-derived basement membrane product?
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MatriMix is a three-dimensional culture matrix composed of collagen, laminin E8 fragments, and hyaluronic acid; which is different from mouse tumor-derived basement membrane products. MatriMix was developed to help progress your research by providing the tools to optimize the extracellular environment suitable for your target cells by changing the type(s), combination(s), and concentration(s) of the collagens and laminin E8 fragments. MatriMix products contain not only basement membrane components, but also stromal collagen, types I and III. If you are not satisfied with your current 3D culture substrates, and are interested in the MatriMix series, please contact us at MatriMix@nippi-inc.co.jp.
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The use and storage of MatriMix is different from that of mouse tumor-derived basement membrane extract products. Please see the MatriMix instruction manual for details.
Can MatriMix be used with cancer patient tissue/cell (PDX/PDC) transplant models?
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Yes, we have confirmed that tumors are formed by transplanting cancer patient derived cell spheroids into mice using MatriMix (PDX/PDSX). We are currently analyzing the details. For more information, please contact us at MatriMix@nippi-inc.co.jp.
It does not gel (gelation is too slow).
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Compared to mouse tumor-derived basement membrane extract (BME) product, MatriMix needs more time to gel. Gelation requires incubation at 37°C for at least 30 minutes. A slight cloudiness is an indication of gelation.
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MatriMix has a mild gelling action even at low temperatures after mixing the three liquids, so it should be used as soon as possible after preparation. MatriMix cannot be frozen. Prepare MatriMix according to the instruction manual before each use.
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Dilution of MatriMix makes it difficult to gel. Whenever possible, especially when culturing in a gel, centrifuge the cell suspension and remove as much of the medium as possible before suspending it with MatriMix soluton. If dilution slows gelation too much, cells will fall to the bottom of the culture vessel.
When trying to prepare an in-gel culture, the cells fall to the bottom of the culture vessel.
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Depending on the cell type and size, the presence or absence of cell aggregates, and the gel volume (height), the cells may fall to the bottom of the culture vessel before gelation. Solutions include (1) pipetting the gel solution into a pre-warmed culture vessel and start incubation immediately, (2) switching to a sandwich culture, or (3) changing the type and/or concentration of collagen.
How can I recover cell spheroids after culture?
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Nippi’s Brightase-C can be used to recover cells by cleaving the collagen. Further trypsin treatment is required to disperse the cell spheroids into individual cells. Please see the protocol for details.
Does the color change when each solution is mixed?
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Solution A (concentrated DMEM, hyaluronic acid/laminin E8) has a grayish blue color. The bluish color comes from the cross-linking reaction between hyaluronic acid and laminin E8.
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Since the concentrated DMEM in solution A contains phenol red, the solution turns pink when mixed with solution B (sodium bicarbonate), and then turns light pink when mixed with solution C (collagen).
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After mixing the 3 solutions, the pH may increase, and the pink color may become darker as time passes. Please use immediately after preparation.
Can MatriMix (511) frozen?
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MatriMix (511) cannot be stored frozen either before or after mixing. Since gelation ability may decrease, store each solution in a refrigerator, and prepare only enough MatriMix for each use.
What components are in MatriMix?
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MatriMix is mainly composed of collagen, hyaluronic acid, and the recombinant human laminin E8 fragments. Other components include DMEM, PBS and sodium bicarbonate.
Does MatriMix contain growth factors?
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No growth factors are added to this product. The main components include: (1) collagen which is derived from animal tissue, (2) hyaluronic acid purified from microorganisms, and (3) laminin E8 purified from CHO-S cell culture supernatant (cells were cultured in a fully defined animal-origin-free media). Each component is purified, so that they are considered to contain little growth factors.
What types of collagen does Nippi offer?
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Nippi can provide various types of collagens from multiple animal origins. For more information, please click here.
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If you purchase Nippi’s reagent grade collagen, you can customize by mixing it with the collagen (solution C) provided with MatriMix. However, the concentration and amount of collagen added may affect gelation. Please contact MatriMix@nippi-inc.co.jp to have collagen expert answer your questions.
Will the gel contract during incubation?
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If there are a large number of stromal cells in the gel, it may contract. The interaction of stromal cells with collagen fibrils causes the gel to shrink.
What is the origin of the laminin E8 fragment?
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A C-terminal fragment of human laminin (E8 fragment) containing a sequence important for integrin binding activity is expressed by CHO-S cells as a recombinant protein. The recombinant laminin E8 proteins are manufactured by Nippi, Inc.