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Advanced BioMatrix/HyStem®-C//GS1005 12.5 mL Kit (Experienced Users Only)
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Product DescriptionHyStem®-C Hydrogel Kits - The starter matrix.HyStem®-C hydrogels provide an excellent starting point for optimizing the matrix for cell culture. HyStem-C is fully chemically-defined and based on three biocompatible components: thiol-modified hyaluronan (Glycosil), thiol-reactive crosslinker, PEGDA (Extralink), and thiol-modified denatured collagen (Gelin-S®). Gelin-S provides basic cell-attachment sites for a wide variety of primary cells and cell lines and is therefore recommended as an ideal substrate for adherent cell types and for cell culture optimization. In some cases, HyStem-C hydrogels can be further enhanced by the addition of ECM proteins to match native signals.FeaturesHydrogels are suitable for culturing primary cells, stem cells and cell lines.Cells can be encapsulated or grown on the hydrogel surface in any format, including culture flasks, 6- to 384-well plates or tissue culture inserts.Hydrogels can be easily customized by the user to possess the desired stiffness and gelation time by manipulating component concentration and mixing ratios.Customizable gelation properties including gelation time and hydrogel stiffness.GelationReconstituted HyStem-C components remain liquid at 15 to 37°C. The hydrogel is formed when the crosslinking agent, Extralink®(PEGDA) is added to a mixture of Glycosil®(thiol-modified hyaluronan) and Gelin-S®(thiol-modified gelatin). Gelation occurs in about twenty minutes after all three components are mixed. No steps depend on low temperatures or low pH. Diluting the components with phosphate-buffered saline (PBS) or cell-culture medium can increase the gelation time.3D Cell Recovery MatrixFor application where cell recovery is critical, the alternative crosslinker PEGSSDA is available for use with all HyStem, HyStem-C and HyStem-HP kits. This crosslinker provides the same advantages offered by Extralink with the additional benefit of containing easily reducible internal bonds. This allows for fast, easy recovery of single cells or clusters from the hydrogel for applications like RNA analysis or flow cytometry instead of slow enzymatic methods that can impact cell viability. Researchers are encouraged to contact us to determine the compatibility of particular cell types or culture systems with PEGSSDA.Directions for UseDownload the HyStem®-Chydrogel kit instructions for:Catalog #GS312 2.5 mL Trial KitCatalog #GS313 7.5 mL KitCatalog #GS1005 12.5 mL KitProduct Q & AWhat size molecules can diffuse through a HyStem hydrogel?Globular particles less than 75 kDa should be able to freely diffuse through a HyStem hydrogel.What is the pH of the reconstituted HyStem components?When reconstituted using DG water, the pH of each HyStem component will be approximately 7.4-7.6.What is the shelf-life of HyStem hydrogel components?One year from the date of receipt, if stored properly.What type of water should be used to reconstitute HyStem?Any sterile, deionized, degassed water can be substituted for reconstitution. However, in order to ensure accurate and predictable dissolution and gelation times, our DG Water is highly recommended, as it is degassed, blanketed in argon, and has undergone validation testing with each HyStem component.Are any cellular attachment sites included in the HyStem hydrogel?Gelin-S provides cellular attachment sites when incorporated in the hydrogel. Gelin-S is thiol-modified, denatured collagen I, derived from either bovine or porcine sources. Gelin-S is included in all HyStem-C and HyStem-HP kits.How does Gelin-S link with the hydrogel?Gelin-S has been thiol-modified in the same manner as the hyaluronan in Glycosil (or Heprasil), so that it covalently crosslinks with the Extralink in the HyStem hydrogels.Can other cell attachment peptides be used in place of Gelin-S?Yes. Peptides that contain a cysteine residue can be used. The cysteine residue must be present for the peptide to be covalently bonded to the hydrogel substrate.Can extracellular membrane (ECM) proteins be used in place of Gelin-S?Yes. ECM proteins, such as laminin, collagen, fibronectin, or vitronectin can be non-covalently incorporated into the hydrogel prior to crosslinking.What is the difference between a HyStem hydrogel and a HyStem sponge?HyStem hydrogels and sponges differ in hydration and homogeneity. HyStem sponges are typically polymerized hydrogels that are subsequently freeze-dried. The resulting sponge is a fibrous, mesh network with pores and niches that enable cells to infiltrate and adhere. A true HyStem hydrogel is an encapsulating liquid that polymerizes around suspended cells in culture.Do HyStem hydrogels change in compliance to stress over time if they are kept in cell culture medium?No. The compliance of the hydrogels is set by the amount of Extralink crosslinker added, the concentration of Glycosil (or Heprasil) and Gelin-S used, and the ratio of Glycosil (or Heprasil) to Gelin-S. Once this chemical structure of the hydrogel is fixed, it is not altered by prolonged exposure to cell culture medium.Can HyStem hydrogels be terminally sterilized?HyStem sponges can be terminally sterilized by E-beam. HyStem hydrogels have not yet been validated for use with E-beam sterilization methods. HyStem hydrogels are not terminally sterilized by gamma irradiation.How can the gelation time be varied when making a HyStem hydrogel?Gelation time is affected by multiple aspects of the gel’s composition.One way to change the gelation time of a hydrogel is to vary the amount of crosslinker used. Gels with a lower amount of Extralink crosslinker will have a longer gelation time than those with a higher amount of crosslinker. Changing the amount of crosslinker will produce slight changes in gelation time.Gelation time can be dramatically changed by varying the Glycosil (or Heprasil) and Gelin-S concentrations. Concentrated solutions of Glycosil (or Heprasil) and Gelin-S will create a solution with a much shorter gelation time. This can easily be done by reconstituting the components in a smaller volume of DG Water. Alternatively, diluting these components in larger volumes of DG Water will dramatically increase the total time to form the hydrogel.Are the cells encapsulated within a HyStem hydrogel easily visible under a microscope?HyStem Hydrogels are virtually transparent and should not interfere with microscopy.Does injection of HyStem hydrogels trigger an immunological or inflammatory response in vivo?HyStem hydrogels may generate mild inflammation as part of the body’s natural healing process in response to injury. HyStem hydrogels do not trigger immune response when used in vivo. (These products are not for human use)What is the mechanism of degredation of HyStem in vivo?HyStem is degraded in vivo by matrix metalloproteinases (collagenases) and hyaluronidases.How can I isolate cells from HyStem hydrogels?Trypsin, Dipase, collagenase, and hyaluronidase have been used to help detach cells from the surface or from within HyStem hydrogels.What is the pore size of HyStem hydrogels?In general, the pore size for HyStem-C and HyStem-HP hydrogels is ~17 nm. Product ApplicationsClick on the title of the desired protocol to learn more:2D Cell Growth on HyStem HydrogelsHyStem 3D Cell Encapsulation for Cell Delivery Applications GuideHyStem 3D Cell Encapsulation in hydrogels using 96-well platesHyStem 3D Cell Encapsulation in hydrogels using TC InsertsEnzyme Digestion of HyStem Hydrogels for Recovery of Encapsulated CellsFluorescent Labeling of HyStem HydrogelsCell Recovery from Surface of HyStem HydrogelsHyStem ECM IncorporationHyStem Gelation Time VariationHyStem Stiffness Variation Protocol for 7.5 mL kitHyStem Stiffness Variation Protocol for 12.5 mL kitProduct ReferencesReferences for HyStem®:Gaetani, R., et al. (2015) Epicardial application of cardiac progenitor cells in a 3D-printed gelatin/hyaluronic acid patch preserves cardiac function after myocardial infarction. Biomaterials 61: 339-348.PMID: 17335875.Prestwich, G.D., et al. (2007) 3-D culture in synthetic extracellular matrices: new tissue models for drug toxicology and cancer drug discovery. Adv Enzyme Regul 47: 196-207.PMID: 17335875.Shu, X.Z., et al. (2006) Synthesis and evaluation of injectable, in situ crosslinkable synthetic extracellular matrices for tissue engineering. J Biomed Mater Res A 79: 901-912.PMID: 16941590.Shu, X.Z., et al. (2003) Disulfide-crosslinked hyaluronan-gelatin hydrogel films: a covalent mimic of the extracellular matrix for in vitro cell growth. Biomaterials 24: 3825-3834.PMID: 12818555.S. Cai, et al. (2005)Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor.Biomaterials, 26, 6054-6067.D. B. Pike, et al. (2006)Heparin-regulated release of growth factors in vitro and angiogenic response in vivo to implanted hyaluronan hydrogels containing VEGF and bFGF.Biomaterials, 27, 5242–5251.G. D. Prestwich, et al. (2007)3-D Culture in Synthetic Extracellular Matrices: New Tissue Models for Drug Toxicology and Cancer Drug Discovery.invited, Adv. Enz. Res., in press (2007).X. Z. Shu, et al, (2006)Synthesis and Evaluation of Injectable, In Situ Crosslinkable Synthetic Extracellular Matrices (sECMs) for Tissue Engineering.J. Biomed Mater. Res. A, 79A(4), 901-912.Shu, X.Z., et al. (2004) In situ crosslinkable hyaluronan hydrogels for tissue engineering. Biomaterials 25: 1339-1348.PMID: 14643608.Mehra, T.D., et al. (2006) Molecular stenting with a crosslinked hyaluronan derivative inhibits collagen gel contraction. J Invest Dermatol 126: 2202-2209.PMID: 16741511.Shu, X.Z., et al. (2004) Attachment and spreading of fibroblasts on an RGD peptide-modified injectable hyaluronan hydrogel. J Biomed Mater Res A 68: 365-375.PMID: 14704979.Ghosh, K., et al. (2007) Cell adaptation to a physiologically relevant ECM mimic with different viscoelastic properties. Biomaterials 28: 671-679.PMID: 17049594.Product Certificate of AnalysisNo result for .Safety and DocumentationCertificate of OriginSafety Data SheetProduct DisclaimerThis product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

美国Advanced BioMatrix3D组织培养、细胞检测和细胞增殖等领域实验解决方案的佼佼者。Advanced BioMatrix在分离、纯化、冻干、细胞培养和蛋白检测、多肽粘附、附着因子、基质硬度和其他3D matrix 产品开发方面有着丰富的经验。Advanced BioMatrix的研发经验和专业知识确保其产品可达到最佳质量,并保证产品之间一致性,方便研究客户使用。

以下为Advanced BioMatrix 3D Matrices 产品竞争优势:
1. 提供高纯度和成分确定的胞外基质;
2. 超过1000余篇文献引用PureCol产品,品质非常均一;
3. 在3D培养基领域可提供最全面的产品线;
4. 唯一可提供特异性刚性有机硅基板的公司(CytoSoft);
5. 唯一可提供可溶性丝纤蛋白的供应商(可运用于多种3D培养);
6. 如果客户首次接触3D胶原凝胶,Advanced BioMatrix还是唯一的预制胶原蛋白(PureCol EZ Gel)供应商。


以下产品为Advanced BioMatrix全球畅销品:
1. PureCol 牛源I型胶原蛋白 3 mg/ml #5005-100ML
2. Nutragen牛源I型胶原蛋白 6 mg/ml #5005-100ML
3. FibriCol 牛源I型胶原蛋白 10 mg/ml #5133-20ML
4. VitroCol 人源I型胶原蛋白 #5007-20ML
5. 弹性蛋白原 #5052-1MG
6. ECM Select Array kit Ultra-36 #5170-1EA
7. CytoSoft(刚性可变的基底,Advanced BioMatrix最新添加产品5190-7EA)
8. 人III型胶原蛋白 #5021-10MG
9. 人IV型胶原蛋白 #5022-5MG
10. Silk Fibroin溶液 #5154-20ML
11. Fibronectin #5080-5MG
12. Vitronectin #5051-0.1MG


Advanced BioMatrix最畅销的产品是PureCol5005-100ML目前已有上千发表的论文使用这款产品。Nutragen 5010fibricol 5133PureCol的物质成分一样,仅仅是浓度不同)浓度越高,胶原蛋白硬度越强。



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