Genetic Engineering & Biotechnology News

OCT1 2012

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TUTORIAL Translational Medicine MSCs were shown to exhibit osteogenic dif- ferentiation following induction, fixation, and staining for alkaline phosphatase activ- ity (Figure 2A). Mineralization by osteocytes was quantified using fluorescence-based staining of hydroxyapatite, and revealed that cells grown on the Fibronectin Mimetic surface exhibit comparable mineralization to cells grown on the human origin matrix (data not shown). MSCs cultured on the Fibronectin surface were also differentiated to adipogenic lineage following induction, fixation, and staining for lipid vacuoles (Fig- ure 2B). Taken together, these results dem- onstrate that human MSCs cultured on the Fibronectin Mimetic surface retain their dif- ferentiation potential. The BD PureCoat ECM Mimetic Colla- gen Type I Peptide surface is prepared using a peptide that contains the GFOGER amino acid motif, which supports the attachment of cells that require a collagen type I coat- ing, including _2-integrin positive cells. This peptide is rationally designed to mimic cell attachment regions present in native collagen type I. The synthetic peptide is covalently im- mobilized on a proprietary surface and pre- sented in a functionally active orientation. This surface is an animal-free alternative to native collagen I or other human-derived ECM surfaces that have been used for kera- tinocyte growth and expansion. Keratinocyte accessibility, proliferation potential, and ease of culture has enabled the use of these cells for cell therapy and regen- erative medicine. Ex vivo expansion of ke- ratinocytes requires either coating a culture vessel with human or animal-derived ECM proteins or culturing cells in growth medium containing bovine serum or animal-derived components. However, the use of animal-or- igin components can raise concerns about in- troducing human-derived or animal-derived pathogens into the culture system and can result in batch-to-batch variability. Our studies demonstrate that human pri- mary keratinocytes can be cultured under serum-free and animal-free conditions on the Collagen I Mimetic surface for multiple pas- sages (Figure 3A) while exhibiting morphol- ogy and growth comparable to cells cultured on native rat tail collagen I or recombinant human collagen I. In addition, keratinocytes retain their functionality after several passages on the Collagen I Mimetic surface as demonstrated by a wound healing assay (Figure 3B). Cells migrated to seal the scratch in the cell mono- layer equally well on all three collagen-based surfaces, resulting in wound healing after 4 hours in culture. Furthermore, the Fibronec- tin Mimetic and Collagen I Mimetic surfaces Marshall Kosovsky, Ph.D. (marshall_ kosovsky@bd.com), is technical support manager at BD Biosciences–Discovery Labware. For further information, please contact Dr. Kosovsky or BD Biosciences– Discovery Labware Technical Support at 877-232-8995 (select prompt 3, then 1) or email at labware@bd.com. have been shown to support the growth and functionality of human endothelial colony forming cells (endothelial progenitor cells) un- der low serum conditions (data not shown). In conclusion, our studies demonstrate that BD PureCoat ECM Mimetic Cul- tureware Fibronectin Peptide supports the xeno-free expansion of human MSCs and maintains their capacity for differentiation. Moreover, BD PureCoat ECM Mimetic Cul- tureware Collagen I Peptide enables the at- tachment, growth, and functionality of hu- man primary keratinocytes in a completely animal-free culture environment. The fully defined and synthetic nature of these novel surfaces greatly reduces the risk of contami- nation from animal-derived pathogens. In addition, these surfaces are manufac- tured in a cGMP-compliant facility, are pre- coated to ensure batch-to-batch consistency, and are available in a variety of scalable for- mats that are room temperature stable for up to 18 months. BD PureCoat ECM Mi- metic Cultureware meets an important need in clinical research by providing animal-free surfaces suitable for the expansion of a vari- ety of cell types. 2012 Vaccine Industry Excellence Awards "Best CMO" Third Year in a Row Connect With Confidence With more than 80 years of parenteral experience, Baxter is a global leader in pre-filled syringe contract manufacturing. BioPharma Solutions' facility in Bloomington, Indiana, USA is one of the largest contract manufacturers in North America, manufacturing over 500 million syringes to date for our pharmaceutical clients. Our offerings include: s ,ARGE ANDSMALL SCALE commercial syringe manufacturing s !BROADRANGEOFDEVELOPMENTSERVICES, including formulation optimization for syringe delivery s !DEDICATEDCLIENTTEAM of cross-functional experts is assigned to each project We provide our clients with confidence of delivery, service, and integrity – because we know that our work is vital to the patients you serve. Let us help you navigate the pathway of success for your molecule. Visit us at: baxterbiopharmasolutions.com Meet with us at: ICSE Europe - Booth #10E25 Baxter is a registered trademark of Baxter International Inc. 920810-00 6/12 Genetic Engineering & Biotechnology News | genengnews.com | October 1, 2012 | 57

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