Product Description
HyStem®-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.Features
- Hydrogels 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 Use
Download the HyStem®-Chydrogel kit instructions for:
Catalog #GS312 2.5 mL Trial Kit
Catalog #GS313 7.5 mL Kit
Catalog #GS1005 12.5 mL Kit
Product Q & A
Globular particles less than 75 kDa should be able to freely diffuse through a HyStem hydrogel.
When reconstituted using DG water, the pH of each HyStem component will be approximately 7.4-7.6.
One year from the date of receipt, if stored properly.
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.
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.
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.
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.
Yes. ECM proteins, such as laminin, collagen, fibronectin, or vitronectin can be non-covalently incorporated into the hydrogel prior to crosslinking.
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.
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.
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.
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.
HyStem Hydrogels are virtually transparent and should not interfere with microscopy.
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)
HyStem is degraded in vivo by matrix metalloproteinases (collagenases) and hyaluronidases.
Trypsin, Dipase, collagenase, and hyaluronidase have been used to help detach cells from the surface or from within HyStem hydrogels.
In general, the pore size for HyStem-C and HyStem-HP hydrogels is ~17 nm.
Product Applications
Click on the title of the desired protocol to learn more:
2D Cell Growth on HyStem Hydrogels
HyStem 3D Cell Encapsulation for Cell Delivery Applications Guide
HyStem 3D Cell Encapsulation in hydrogels using 96-well plates
HyStem 3D Cell Encapsulation in hydrogels using TC Inserts
Enzyme Digestion of HyStem Hydrogels for Recovery of Encapsulated Cells
Fluorescent Labeling of HyStem Hydrogels
Cell Recovery from Surface of HyStem Hydrogels
HyStem ECM Incorporation
HyStem Gelation Time Variation
HyStem Stiffness Variation Protocol for 7.5 mL kit
HyStem Stiffness Variation Protocol for 12.5 mL kit
Product References
References 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 Analysis
Safety and Documentation
Certificate of Origin
Safety Data Sheet
Product Disclaimer
This 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AdvancedBioMatrix（简称ABM） www.advancedbiomatrix.comAdvancedBioMatrix（简称ABM）是美国一家著名的生物公司，获得了AllerganInc的授权(Allergan用25年时间不断完善胶原蛋白相关的产品的生产工艺)，将Allergan的专业和技术用于蛋白生产与检测，致力于为组织工程、细胞分析及细胞增殖等研究领域提供优质稳定的产品。AdvancedBioMatrix不断丰富已有产品线，目前可为三维细胞培养提供各种胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、不同粘度与分子量的透明质酸以及低代成纤维细胞等。在美国全部产品授权Sigma销售。AdvancedBioMatrix是组织培养，细胞分析和细胞增殖三维（3D）应用的生命科学领域的领导者。我们的产品被公认为纯度，功能性和一致性的标准。我们在生产，分离，纯化，冷冻干燥，细胞培养和蛋白质测试，粘附肽，附着因子，底物刚性和其他3D矩阵产品方面拥有丰富的专业知识。我们的专业技术和知识正在被用来确保我们的产品质量最高，批次之间一致且易于为我们的研究客户使用。
美国AdvancedBioMatrix是3D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。AdvancedBioMatrix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3Dmatrix 产品开发方面有着丰富的经验。AdvancedBioMatrix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AdvancedBioMatrix3DMatrices 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，AdvancedBioMatrix还是唯一的预制胶原蛋白(PureColEZGel)供应商；
以下产品为AdvancedBioMatrix全球畅销品：1.PureCol 牛源I型胶原蛋白 3mg/ml#5005-100ML2.Nutragen牛源I型胶原蛋白 6mg/ml#5010-50ML3.FibriCol 牛源I型胶原蛋白 10mg/ml#5133-20ML4.VitroCol 人源I型胶原蛋白 #5007-20ML5. 弹性蛋白原 #5052-1MG6.ECMSelectArraykitUltra-36#5170-1EA7.CytoSoft（刚性可变的基底，AdvancedBioMatrix最新添加产品5190-7EA）8. 人III型胶原蛋白 #5021-10MG9. 人IV型胶原蛋白 #5022-5MG10.SilkFibroin溶液 #5154-20ML11.Fibronectin#5080-5MG12.Vitronectin#5051-0.1MG
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首先要什么有什么的，你得好好考虑一下。
2、看生产地址
根本没有生产地址，我们知道做实验做产品需要很多的仪器、试剂、耗材，没有人相信一间简单的屋子可以生产各种样的试剂盒。
3、看产品包装
没有任何的生产地址、联系方式等信息，这种产品有问题了连个投诉的地方都没有。
4、看公司网站
有些打着国外原装旗号，整个公司网站为英文页面，实际注册IP地址在中国。如果写着国外的地址，让你国外的朋友实地去看一下！
5、做交叉验证
拿对方提供的几个种类的试剂盒，把里面的关键组份相互替换做做实验，如果交叉严重，只能说明是一种原料生产的试剂盒贴了不同的标签。
6、看价格
价格低得离谱，却打着进口大公司原料分装，核算成本，这种低得离谱的价格是连原料都买不起的。
现在国内最差也是用3代试剂，有些地方会用4代试剂。
4代试剂（检查抗原+抗体）——窗口期为4周。因为抗原于3-4周达到复制的峰值，此时通过4代试剂检查，如果感染了HIV，抗原/抗体至少有一个为阳性，如果都是阴就排除了。
3代试剂（只查抗体）——窗口期为6周。
以上为理论分析+临床经验的结果，可以说是99.9%的准确度。
但是目前FDA、CDC和试剂生产商统一达成的共识，也就是针对普通人，最保守的窗口期是3个月。无论什么试剂，3个月都100%排除。
ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性，酶标记的抗原或抗体既保留其免疫学活性，又保留酶的活性。在测定时，受检标本（测定其中的抗体或抗原）与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体，也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后，底物被酶催化成为有色产物，产物的量与标本中受检物质的量直接相关，故可根据呈色的深浅进行定性或定量分析。由于酶的催化效率很高，间接地放大了免疫反应的结果，使测定方法达到很高的敏感度。
1．取出试剂盒室温平衡30min，取出血样放至室温。
2．配标准品：取150uL标准品加入150uL标准品稀释液稀释，依次稀释5次。
3．加样：分别于各反应孔中加入标准品50uL，样品40UL，标准品做复孔，样品做3孔。
4．分别于样品孔中加入10UL抗体。
5．标准品和样品孔中分别加入50uL链酶亲和素-HRP，盖上封板膜,轻轻震荡混匀，37℃温育60min。
6．配洗涤液：将30倍浓缩洗涤液用蒸馏水30倍稀释后备用。
7．洗涤：小心揭开封板膜，弃去液体，甩干，每孔加200uL洗涤液，静置30s后弃去，如此重复5次，拍干。
8．显色：每孔先加入显色剂A 50uL，再加显色剂B 50uL，轻轻震荡混匀，37℃避光显色6min。
9． 终止：每孔加入终止液50uL，终止反应（此时蓝色立即转为黄色）。
10．测定：以空白孔调零，450nm波长依序测量各孔的吸光度。测定应在加终止液10min之内进行。
11．保存结果，收拾桌面。
12．分析处理数据
注意事项
1. 取出板条前恢复到室温后再打开外包装袋，实验中不用的板条立即放回包装中，密闭封口，其余不用试剂应盖好。
2. 实验操作中请使用一次性的吸头，避免交叉污染。
3．实验板孔加入试剂的顺序应一致，以保证所有反应孔的孵育时间一致。
4．洗涤过程中反应孔中残留的洗涤液应在滤纸上充分拍干，勿将滤纸直接放入反应孔中吸水。
5．试剂盒内试剂请在保质期内使用，不同批号试剂不要混用。
6．1000pg/ml以上的结果为非线性的，根据此标准曲线无法得到精确的结果。大于1000pg/ml 的样品应以标准稀释缓冲液稀释后重做。在结果分析时，结合考虑相应的稀释度。
