BD™ PuraMatrix™ Peptide Hydrogel Guidelines for Use - 3DM Inc.

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TABLE OF CONTENTS Intended Use........................................................................................................................................ 2 Materials Provided............................................................................................................................... 3 Supplemental Materials (not supplied) ................................................................................................ 3 Precautions ......................................................................................................................................... 3 General Use……………………………………………………….............................................................. 3 Procedures for Use........................................................................................................................... 5-14 A) Surface Plating of Adherent Cells in Cell Culture Plates…………………………………………....... 5 B) Surface Plating of Adherent Cells in Cell Culture Inserts………………………………………….... 5-6 C) ECM Preparation for Supplementation of BD PuraMatrix Peptide Hydrogel………………….. 7 D) 3D Cell Encapsulation in Cell Culture Plates…………………………………………………………… 8 E) 3D Cell Encapsulation in Cell Culture Inserts …………………………………..…………………… 8-11 F) Cell Drop Culture Protocol.............................................................................................................. 12 G) Cell Recovery for Sub-Culturing or Biochemical Analyses…………………………………………… 13 H) Fluorescent Staining of Samples Cultured with BD PuraMatrix Peptide Hydrogel……………… 13-14 I) Tips for In Vivo Use in Animals………………………………………………………………………...…. 15 J) Recommended Plating Volumes for Multi-well Plates & Inserts……………………………………... 16 Stability................................................................................................................................................ 16 Technical Service ............................................................................................................................... 16 2
INTENDED USE Synthetic scaffolds for defined cellular microenvironments BD PuraMatrix Peptide Hydrogel is a synthetic matrix that is used to create defined three dimensional (3D) microenvironments for a variety of cell culture experiments. To achieve optimal cell growth and differentiation, it is necessary to determine the appropriate mixture of this material and bioactive molecules (e.g., growth factors, extracellular matrix (ECM) proteins, and/or other molecules). BD PuraMatrix Peptide Hydrogel consists of standard amino acids (1% w/v) and 99% water. Under physiological conditions, the peptide component of BD PuraMatrix self-assembles into a 3D hydrogel that exhibits a nanometer scale fibrous structure with an average pore size of 50-200 nm. The hydrogel is readily formed in a culture dish, plate, or cell culture insert. Advantages of 3D cell structures with the ease of 2D cell plating BD PuraMatrix often provides the benefits of 3D cell culture while allowing more straightforward 2D plating. Many cell types form appropriate 3D morphologies when plated on the surface of BD PuraMatrix, which simplifies the work required to develop 3D in vitro model systems compared with encapsulation or sandwich cultures in other gels. As research has already shown that cells forming 3D structures often resemble their in vivo counterparts more closely than when grown in two dimensions, BD PuraMatrix allows defined microenvironments to better recapitulate the in vivo milieu, which should improve the predictive quality of hits obtained from assays conducted in a high throughput context. Encapsulates cells & growth factors, both in vitro and in vivo The hydrogel has been shown to support the differentiation of hepatocyte progenitor cells 1 , rat pheochromocytoma cells (PC12) 2 and hippocampal neurons 3 , as well as enabling endothelial cell tubulogenesis 4 . Studies have also demonstrated that BD PuraMatrix supports the attachment of a variety of primary (e.g., neuronal, fibroblast, keratinocyte) and transformed (e.g., MG-63, SH-SY5Y, HEK293, NIH3T3) cell types 5 . Other potential applications include stem cell proliferation, tumor cell migration and in vivo analyses of tissue regeneration. BD PuraMatrix is biocompatible, resorbable, and devoid of animalderived material and pathogens. For in vivo studies in animals, the soluble material can be injected and will subsequently form a 3D hydrogel upon contact with the physiological environment. REFERENCES 1. Semino, CE, et al. (2003) Differentiation 71:262-270. 2. Holmes, TC, et al. (2000) PNAS USA 97:6728-6733. 3. Semino, CE, et al. (2004) Tissue Engineering 10:643-655. 4. Narmoneva, D, et al. (2005) Biomaterials 26:4837-4846 5. Zhang, S, et al. (1995) Biomaterials 16:1385-1393. For additional references, go to http://www.bdbiosciences.com/discovery_labware/products/display_product.php?keyID=179 THIS PRODUCT IS FOR RESEARCH USE ONLY; NOT FOR CLINICAL, DIAGNOSTIC OR THERAPEUTIC USE. NOT FOR USE IN HUMANS. 3
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BD™ PuraMatrix™ Peptide Hydrogel Guidelines for Use - 3DM Inc.

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