Animal-derived Materials Research Articles

Bodily Traces: Animal Matter, Historical Books and the ‘Lifelessness’ of Writing

Throughout the medieval and early modern periods in Europe, animal-derived materials were routinely used in book production, and thousands of animals thus left their bodily traces in and on supposedly human cultural artefacts. Acknowledging their presence requires us to rethink how we conceive of life and presence in relation to writing and of animals in relation to textuality. The animals whose remains were turned into books not only embody meaning but their bodies became meaning in the shape of text. This article sets out to follow the material traces of these animals rendered invisible centuries ago to draw attention to the tangible animality of our literary heritage. By raising consciousness for the essential, yet often neglected role other animals played in its creation, my research troubles traditional animal-human and nature-culture binaries, calling for a more nuanced appreciation of animal lives in- and outside (human) texts.

Miniaturized droplet microarray platform enables maintenance of human induced pluripotent stem cell pluripotency.

The capacity of human induced pluripotent stem cells (hiPSCs) for indefinite self-renewal warrants their application in disease modeling, drug discovery, toxicity assays and efficacy screening. However, their poor proliferation ability, inability to adhere to surfaces without Matrigel coating and tendency to spontaneously differentiate in vitro hinder the application of hiPSCs in these fields. Here we study the ability to culture hiPSCs inside 200 ​nL droplets on the droplet microarray (DMA) platform. We demonstrate that (1) hiPSCs can attach to the Matrigel (MG)-free surface of DMA and show good viability after 24 h culture; (2) hiPSC do not spontaneously differentiate when cultured on the MG-free surface of DMAs; (3) culturing of hiPSCs in 200 ​nL as compared to 2 ​mL culture leads to higher expression of the Nanog pluripotency marker. Overall, the results demonstrate the possibility to culture undifferentiated hiPSCs in 200 ​nL droplets on DMA, thereby opening the possibility for high-throughput screenings of hiPSCs with various factors without compromising the results through the involvement of animal-derived materials, such as Matrigel.

Bone Graft Biomineral Complex Coderived from Marine Biocalcification and Biosilicification.

Bone graft materials have been mainly developed based on inorganic materials, including calcium phosphate. However, these graft materials usually act as osteoconductive rather than osteoinductive scaffolds. To improve bone reconstruction, a combination of several materials has been proposed. However, there are still no alternatives that can completely replace the existing animal-derived bone graft materials. In this work, a marine-inspired biomineral complex was suggested as a potential bone graft material. The proposed biosilicified coccolithophore-derived coccoliths using bioengineered mussel adhesive proteins show osteopromotive ability through the synergistic effects of osteoconductivity from calcium carbonate and osteoinductivity from silica. Its possibility of use as a bone substitute was determined by evaluating the in vitro osteogenic behaviors of multipotent mesenchymal stem cells and in vivo bone regeneration in a rat calvarial defect model. Therefore, the marine-inspired biomineral complex developed in this study could be successfully used for bone tissue engineering.

Mobile Elements Harboring Heavy Metal and Bacitracin Resistance Genes Are Common among Listeria monocytogenes Strains Persisting on Dairy Farms.

ABSTRACTListeria monocytogenes is a foodborne pathogen and a resilient environmental saprophyte. Dairy farms are a reservoir of L. monocytogenes, and strains can persist on farms for years. Here, we sequenced the genomes of 250 L. monocytogenes isolates to investigate the persistence and mobile genetic elements (MGEs) of Listeria strains inhabiting dairy farms. We performed a single-nucleotide polymorphism (SNP)-based phylogenomic analysis to identify 14 monophyletic clades of L. monocytogenes persistent on the farms for ≥6 months. We found that prophages and other mobile genetic elements were, on average, more numerous among isolates in persistent than nonpersistent clades, and we demonstrated that resistance genes against bacitracin, arsenic, and cadmium were significantly more prevalent among isolates in persistent than nonpersistent clades. We identified a diversity of mobile elements among the 250 farm isolates, including three novel plasmids, three novel transposons, and a novel prophage harboring cadmium resistance genes. Several of the mobile elements we identified in Listeria were identical to the mobile elements of enterococci, which is indicative of recent transfer between these genera. Through a genome-wide association study, we discovered that three putative defense systems against invading prophages and plasmids were negatively associated with persistence on farms. Our findings suggest that mobile elements support the persistence of L. monocytogenes on dairy farms and that L. monocytogenes inhabiting the agroecosystem is a potential reservoir of mobile elements that may spread to the food industry.IMPORTANCE Animal-derived raw materials are an important source of L. monocytogenes in the food industry. Knowledge of the factors contributing to the pathogen’s transmission and persistence on farms is essential for designing effective strategies against the spread of the pathogen from farm to fork. An increasing body of evidence suggests that mobile genetic elements support the adaptation and persistence of L. monocytogenes in the food industry, as these elements contribute to the dissemination of genes encoding favorable phenotypes, such as resilience against biocides. Understanding of the role of farms as a potential reservoir of these elements is needed for managing the transmission of mobile elements across the food chain. Because L. monocytogenes coinhabits the farm ecosystem with a diversity of other bacterial species, it is important to assess the degree to which genetic elements are exchanged between Listeria and other species, as such exchanges may contribute to the rise of novel resistance phenotypes.

Switching Kogenate FS® to Kovaltry® for the Treatment of Severe Haemophilia A Patients: A Successful Single Center Experience

Background: Haemophilia A is an X-linked hereditary disorder caused by deficient levels of the coagulation protein factor VIII (FVIII). Standard therapy for haemophilia A comprises replacement of FVIII with plasma-derived or recombinant FVIII concentrate, which can be administered as on-demand or prophylaxis regimen. The most challenging complication of replacement therapy in hemophilia A is the occurrence of alloantibodies against infused factor FVIII, thus predisposing the patients to an increased morbidity and disability. Historically, first FVIII was isolated from cryoprecipitated plasma concentrates. Thirty years ago, after the cloning of F8 gene, therapy for hemophilia A patients underwent a new exciting era that developed technological strategies aimed to improve haemostatic efficacy and safety of the replacement factor. In this line, a new recombinant FVIII, BAY 81-8973 (octocog alfa; Kovaltry®, Bayer) has recently been introduced onto pharmaceutical market for treatment of these patients. Kovaltry® is an unmodified, full-length, recombinant FVIII concentrate with the same amino acid sequence as Kogenate FS® (Bayer), but produced with innovative manufacturing technologies that include production without addition of human- or animal-derived raw materials from purification or formulation processes, inclusion of a higher degree of sialic acid capping of N-terminal glycans on the molecular surface, coexpression of the human chaperone protein heat shock protein 70, and a 20-nm nanofiltration step to remove small non-enveloped viruses and potential protein aggregates. On this basis we considered beneficial for our patients changing Kogenate FS® therapy for Kovaltry®.Objective: We aimed to demonstrate that switching from Kogenate FS® to Kovaltry® could be easily performed without increasing the risk of bleeding or FVIII consumption in severe haemophilia A patients.Methods: Sixty two adult and pediatric patients with severe haemophilia A, without inhibitors, treated with Kogenate FS® either on on-demand treatment (n=13) or on prophylaxis regimen (n=49), were recruited. On November 2016, we began to switch Kogenate FS® treatment to Kovaltry® in these patients.The trend of prophylaxis with Kovaltry® was evaluated by determining mean injection frequency (per week), mean dose of each injection (IU/kg), mean total dose injected/week (IU), baseline and post-infusion levels of FVIII and annual bleeding rate (ABR). Same data for Kogenate FS® were collected retrospectively from clinical histories of patients.For determining baseline and post-infusion levels of Kovaltry®, whole blood was collected in 3.2% sodium citrate tubes (BD Vacutainer, Becton Dickinson, San José, CA, USA) at pre-dosing and 30 minutes after Kovaltry® infusion. Plasma FVIII:C was measured by chromogenic assay.Results: Switching from Kogenate FS® to Kovaltry® caused no changes, except in two patients, either in mean injection frequency or mean dose of each injection or mean total dose injected/week or baseline and post-infusion levels of FVIII or annual bleeding rate. Increment observed in mean total dose injected/week in two of the pediatric patients was due to an increased physical activity (patient 2) and to weight gain (patient 8). For sake of example, Table 1 shows these variables for 10 pediatric patients under prophylactic treatment.None of the patients treated with Kovaltry® developed inhibitors.Conclusion: Changing severe haemophilia A treatment from Kogenate FS® to Kovaltry® can be performed easily and did not pose any risk to the patient since Kovaltry® proved to be equally effective than Kogenate FS® to prevent bleeding episodes and did not induce the occurrence of inhibitors. [Display omitted] DisclosuresÁlvarez-Roman:Novo Nordisk: Consultancy; Shire: Consultancy. Jimenez-Yuste:Novo Nordisk: Consultancy, Honoraria, Research Funding; Roche: Consultancy.

Role of Phage Capsid in the Resistance to UV-C Radiations.

The conformational variation of the viral capsid structure plays an essential role both for the environmental resistance and acid nuclear release during cellular infection. The aim of this study was to evaluate how capsid rearrangement in engineered phages of M13 protects viral DNA and peptide bonds from damage induced by UV-C radiation. From in silico 3D modelling analysis, two M13 engineered phage clones, namely P9b and 12III1, were chosen for (i) chemical features of amino acids sequences, (ii) rearrangements in the secondary structure of their pVIII proteins and (iii) in turn the interactions involved in phage capsid. Then, their resistance to UV-C radiation and hydrogen peroxide (H2O2) was compared to M13 wild-type vector (pC89) without peptide insert. Results showed that both the phage clones acquired an advantage against direct radiation damage, due to a reorganization of interactions in the capsid for an increase of H-bond and steric interactions. However, only P9b had an increase in resistance against H2O2. These results could help to understand the molecular mechanisms involved in the stability of new virus variants, also providing quick and necessary information to develop effective protocols in the virus inactivation for human activities, such as safety foods and animal-derived materials.

Insulation of Common Chaffinch Fringilla coelebs Nests is Largely Driven by Animal-Derived Materials in the Cup Lining

Incubation of eggs and nestlings is energetically costly and may be affected by the amount of thermal insulation the surrounding nest provides. In many small passerine species, animal-derived materials, i.e., feathers and hair, are often used in the nest lining where, presumably, their lower thermal conductance offers better insulation. A previous study that involved partial deconstruction of thrush nests revealed that different parts of the nest confer different levels of insulation. The present study examined the insulatory values of Common Chaffinch Fringilla coelebs nests because of the high proportion of animal-derived materials in their cup lining but not the outer nest. Insulatory values and cooling rates of temperature loggers inside the cup were determined for whole nests and then only the cup lining. There were no significant relationships between nest measurements and measures of thermal insulation. However, removal of the outer nest wall reduced the insulatory value by around 10% despite the cup lining being less than half the thickness of the complete nest wall and the cup lining being only a third of the total nest mass. Differential placement of animal-derived materials within the cup lining of Common Chaffinch nests means that the birds seem to be able to confer a high level of insulation without expending too much energy searching for a large mass of particular nest materials.

Multimodality Imaging of Foreign Bodies: New Insights into Old Challenges.

Traumatic wounds and lacerations are a common reason for patients to present to emergency departments, with retained foreign bodies (FBs) accounting for 7%-15% of cases, particularly those involving the extremities. These retained materials result in a granulomatous tissue response known as an FB reaction, a pathologic attempt to isolate the FB from the host. The most common FB materials are glass, metal, and wood, but other compositions can also be found, such as plastic and animal-derived materials. Clinical history, physical examination, and wound exploration are essential in investigation of retained material but are not sufficient to exclude an FB, and additional investigation is required. Imaging evaluation is a useful tool to help depict and locate an FB, assess possible complications, and guide removal. Conventional radiography, the first-line method in this scenario, is a widely available low-cost depiction method that has good sensitivity for depicting FBs. If the retained material is not depicted at conventional radiography, US can be performed. US is highly sensitive in depicting both radiolucent and radiopaque FBs in superficial locations. For deeper objects, CT may be necessary. MRI is the best imaging modality to delineate local soft-tissue and osseous complications. Retained FBs can result in early and delayed complications, with infection being the most frequent complication. To avoid preventable morbidities related to FBs, radiologists should be familiar with imaging findings and provide essential information to help the attending physician treat each patient. Online supplemental material is available for this article. ©RSNA, 2020.

Poly(POG)n loaded with recombinant human bone morphogenetic protein-2 accelerates new bone formation in a critical-sized bone defect mouse model

BackgroundDelivery of bone morphogenetic protein-2 (BMP-2) via animal-derived absorbable collagen materials is used for the treatment of large bone defects. However, the administration of bovine proteins to humans is associated with the risk of zoonotic complications. We therefore examined the effect of combining BMP-2 with collagen-like peptides, poly(POG)n, in a critical-sized bone defect mouse model.MethodsA 2-mm critical-sized bone defect was created in the femur of 9-week-old male C57/BL6J mice. Mice were randomly allocated into one of four treatment groups (n = 6 each): control (no treatment), poly(POG)n only, 0.2 μg, or 2.0 μg BMP-2 with poly(POG)n. New bone formation was monitored using soft X-ray radiographs, and bone formation at the bone defect site was examined using micro-computed tomography and histological examination at 4 weeks after surgery.ResultsAdministration of 2.0 μg of BMP-2 with poly(POG)n promoted new bone formation and resulted in greater bone volume and bone mineral content than that observed in the control group and successfully achieved consolidation. In contrast, bone formation in all other groups was scarce.ConclusionsOur findings suggest the potential of BMP-2 with poly(POG)n as a material, free from animal-derived collagen, for the treatment of large bone defects.

De-risking the immunotherapy supply chain: development of virus inactivated human AB serum for immune cell expansion

Background & Aim FBS and Calf bovine serum from USDA-approved animals are used in a broad range of cell culture applications; they have been demonstrated to be safe and effective growth stimuli. However, the use of xenogenic serum always implies a certain degree of risk associated with a pathogen-driven immune response or the unwanted incorporation of foreign proteins into human primary cells, such as stem cells, T cells and NK cells. These risks have driven demand for xenogeneic-free culture media for clinical handling of these cells. Akron Biotech has developed several off-the-shelf AB serum options that allow cell therapy developers to avoid the risk inherent in xenogenic sera. Akron's latest development is the development of a virus inactivated human AB serum that incorporates solvents/detergents to improve the safety of this critical supplement for immunotherapy development without compromising its efficacy in promoting cell growth. Methods, Results & Conclusion Methods Process development schematic figure is attached. The biochemical profile and quality of AB serum (native components #1, #2, and #3 as well as additional components) batches were analyzed. Their ability to support cell growth after two days cell culture was evaluated by crystal violet assay and cell proliferation assay. Cell lines: 293T, Jurkat T cell and PBMC-derived T cells. Results The three processed serum sources (#1, #2, #3) were demonstrated to effectively promote cell growth. The three cell lines performed similarly or better than converted AB serum. Overall, our results indicate that virus inactivated AB serum is a safe and efficient material for inducing cell proliferation that does not negatively impact cell quality. Given the wellunderstood problems associated with using FBS and other animal-derived materials, we believe that our virus inactivated AB serum, developed through extensive process development, is the safest alternative for the proliferation of cells critical to immunotherapy development and clinical application.

Feasibility of defined, animal component free, protein free, salt base freezing solution composed of 5% DMSO for cryopreservation of HMSC toward clinical applications

Background & Aim hMSC are a critical raw material for cell-based therapies and usually a large number of cells are required for clinical applications. Cryopreservation is currently the only method to preserve cells with maintained functional properties and genetic stability for long term storage. To date, a common practice is use of homebrew cryo-medium that is composed of animal-derived raw materials and low Mw Cryoprotective agents (CPAs), usually 10-20% DMSO. Alternatively, animal component free (ACF) freezing solutions were developed for clinical applications, but they are still based on culture medium and are composed of 10% DMSO (e.g. NutriFreez™ D10). In addition, a few products were developed with a reduced concentration of DMSO or even without DMSO. However, these products appear not to be optimal for hMSC. Moreover, most of the commercially available DMSO-free products are actually composed of other potentially toxic permeable CPAs (e.g. Ethylene Glycol, EG). Exposure of cells to these materials can impact the quality, safety and efficacy of cell-based therapeutic product. Facing strict regulatory requirements, the development of defined, ACF, salt base freezing solution with reduced concentration of DMSO is essential. Methods, Results & Conclusion The current study presents the feasibility of a novel defined, ACF, protein free, salt base freezing solution composed of 5% DMSO for cryopreservation of hMSC from various sources. Results show that the novel cryopreservation solution efficiently maintains high cell yield and viability, and supports the recovery of hMSC while maintaining the normal hMSC features: typical fibroblast-like cell morphology, phenotypic surface marker profile, differentiation capability as well as self-renewal potential. Clinically accepted, cryopreservation solution for hMSC holds a unique opportunity to facilitate the translation of these cells to cellular therapy applications.

Material Type and Position Determines the Insulative Properties of Simulated Nest Walls

Incubation in birds takes place within a nest that is often assumed to confer a degree of thermal insulation. The range, amounts and organisation of materials used to construct nest walls hampers our understanding of the degree to which they provide insulation during incubation. This experimental study used temperature loggers in a model system to test the insulative properties of materials extracted from bird nests to determine: 1) whether differences existed in terms of insulation, and 2) if the position of a material mattered when two materials were tested in combination. Animal-derived materials offered better insulation than plant-derived materials, whether tested singly or in combination. Halving the mass of each material did not affect insulation conferred by the material proximal to the temperature logger. Differing thermal conductivities of the materials in contact with the temperature logger may explain these results. If a bird strategically places an animal-derived material only into a nest cup lining then it may be sufficient to provide good insulation for the whole nest. More research is needed to generate thermal conductivity data for commonly used nest materials to test this idea more rigorously in finite element heat transfer models.—Deeming, D.C., Griffiths, J.D. & Biddle, L.E. (2020). Material type and position determines the insulative properties of simulated nest walls. Ardeola, 67: 127-136.

Identification of a distinct lineage of aviadenovirus from crane feces.

Viruses are believed to be ubiquitous; however, the diversity of viruses is largely unknown because of the bias of previous research toward pathogenic viruses. Deep sequencing is a promising and unbiased approach to detect viruses from animal-derived materials. Although cranes are known to be infected by several viruses such as influenza A viruses, previous studies targeted limited species of viruses, and thus viruses that infect cranes have not been extensively studied. In this study, we collected crane fecal samples in the Izumi plain in Japan, which is an overwintering site for cranes, and performed metagenomic shotgun sequencing analyses. We detected aviadenovirus-like sequences in the fecal samples and tentatively named the discovered virus crane-associated adenovirus 1 (CrAdV-1). We determined that our sequence accounted for approximately three-fourths of the estimated CrAdV-1 genome size (33,245bp). The GC content of CrAdV-1 genome is 34.1%, which is considerably lower than that of other aviadenoviruses. Phylogenetic analyses revealed that CrAdV-1 clusters with members of the genus Aviadenovirus, but is distantly related to the previously identified aviadenoviruses. The protein sequence divergence between the DNA polymerase of CrAdV-1 and those of other aviadenoviruses is 45.2-46.8%. Based on these results and the species demarcation for the family Adenoviridae, we propose that CrAdV-1 be classified as a new species in the genus Aviadenovirus. Results of this study contribute to a deeper understanding of the diversity and evolution of viruses and provide additional information on viruses that infect cranes, which might lead to protection of the endangered species of cranes.

SOLUTION PLASMA PROCESSING OF NATURAL POLYMER-BASED MATERIALS

Non-equilibrium low-temperature atmospheric pressure plasma is a highly reactive medium that allows for the ecofriendly modification of nature-derived materials. This work reviews the field of liquid phase plasma modification of polymer-based materials derived from natural sources. Types of solution plasma systems are considered with respect to the efficiency of the generation of active species, their transport to and interaction with macromolecules. Tunable modification of plants (birch, willow, bamboo and vine shoots; barley, rice and rapeseed straw; perennial grasses), algae, plant-derived by-products and wastes (sawdust, bast fibers, nutshells, seeds, peels, press-cake and pomace), plant-derived polymers (cellulose, hemicellulose, lignin, starch, sodium alginate) and animal-derived polymer-based materials (chitosan, collagen, gelatin) by solution plasma treatment is compared to conventional wet chemistry methods. Synthesis of nano-biocomposites with advanced functionalities is also considered.
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Elicited and pre-existing anti-Neu5Gc antibodies differentially affect human endothelial cells transcriptome.

Humans cannot synthesize N-glycolylneuraminic acid (Neu5Gc) but dietary Neu5Gc can be absorbed and deposited on endothelial cells (ECs) and diet-induced anti-Neu5Gc antibodies (Abs) develop early in human life. While the interaction of Neu5Gc and diet-induced anti-Neu5Gc Abs occurs in all normal individuals, endothelium activation by elicited anti-Neu5Gc Abs following a challenge with animal-derived materials, such as following xenotransplantation, had been postulated. Ten primary human EC preparations were cultured with affinity-purified anti-Neu5Gc Abs from human sera obtained before or after exposure to Neu5Gc-glycosylated rabbit IgGs (elicited Abs). RNAs of each EC preparation stimulated in various conditions by purified Abs were exhaustively sequenced. EC transcriptomic patterns induced by elicited anti-Neu5Gc Abs, compared with pre-existing ones, were analyzed. qPCR, cytokines/chemokines release, and apoptosis were tested on some EC preparations. The data showed that anti-Neu5Gc Absinduced 967 differentially expressed (DE) genes. Most DE genes are shared following EC activation by pre-existing or anti-human T-cell globulin (ATG)-elicited anti-Neu5Gc Abs. Compared with pre-existing anti-Neu5GcAbs, which are normal component of ECs environment, elicited anti-Neu5Gc Abs down-regulated 66 genes, including master genes of EC function. Furthermore, elicited anti-Neu5Gc Abs combined with complement-containing serum down-regulated most transcripts mobilized by serum alone. Both types of anti-Neu5Gc Abs-induced a dose- and complement-dependent release of selected cytokines and chemokines. Altogether, these data show that, compared with pre-existing anti-Neu5Gc Abs, ATG-elicited anti-Neu5Gc Abs specifically modulate genes related to cytokine responses, MAPkinase cascades, chemotaxis, and integrins and do not skew the EC transcriptome toward a pro-inflammatory profile in vitro.

Traditional medicinal animal use by Xhosa and Sotho communities in the Western Cape Province, South Africa

BackgroundThe use of animals and animal-derived materials in traditional medicine constitutes an important part of the belief systems of indigenous African cultures. It is believed to be rapidly expanding in South Africa, where traditional healers are estimated to outnumber western doctors by 2000:1 in some areas, with an overall clientele consisting of 60–80% of South African citizens. Despite concerns about the impact of the trade in traditional medicine on biodiversity, there has been only limited research on this topic in South Africa.MethodsTraditional Xhosa and Sotho healers operating from impoverished, rural communities in the Boland Region of the Western Cape Province were consulted to provide a comprehensive inventory of the number and frequency of animals used and sold. Species richness estimators, diversity indices, and a relative cultural importance (RCI) index were used to highlight species of concern and assess market dynamics.ResultsA total of 26 broad use categories for 12 types of animal parts or products from 71 species or morphospecies were recorded. The most commonly sold items were skin pieces, oil or fat, and bones. Results showed that leopard, chacma baboon, Cape porcupine, monitor lizard species, puff adder, African rock python, and black-backed jackal were the species most used in the traditional medicinal trade.ConclusionsThis study extends existing knowledge on the trade of animals in South African healing practices and provides the first attempt in the Western Cape to quantify wildlife use for cultural traditions. The results have relevance for setting conservation priorities and may assist in effective policy development inclusive of ecological sustainability priorities, as well as cultural demands.

Long‐term culture of human pluripotent stem cells on coverslips coated with peptide‐poly(styrene‐co‐vinyl benzoic acid) copolymer

Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have the potential to grow indefinitely (self‐renewal) in culture and differentiate into any cell type in adult human body. The hESCs and hiPSCs represent important resources for regenerative medicine and for disease modeling. Therefore, growing stem cells in undifferentiated state is important. The human pluripotent stem cells are expanded traditionally using either mitotically inactivated mouse embryonic fibroblasts (feeder cells) or Matrigel (MG). Both feeder cells and MG are prepared using mouse tissues that have several disadvantages. These manufactured expensive biological materials that have limited scalability also have high batch‐to‐batch variability. In addition, the animal‐derived (xenogenic) materials have potential to spread inter‐species pathogens. Thus, in order to replace these animal based biological materials, great efforts have been invested to develop synthetic materials that could be used for long‐term pluripotent stem cell growth and proliferation. As a consequence, materials such as poly [2‐(methacryloyloxy) ethyl dimethyl‐(3‐sulfopropyl) ammonium hydroxide] (PMEDSAH) and UV treated peptide‐acrylate materials have been used to culture hESCs. Since these synthetic materials are polymerized directly on the surface of cell culture dish, its application in broad sector and large‐scale expansion of stem cells for downstream applications has limitation. Thus, in the present study, we used a copolymer system containing poly (vinyl benzoic acid‐co‐styrene) (PVBA‐St), which could be coated onto various cell culture surfaces, such as glass, polystyrene cell culture plate etc. We synthesized PVBA‐St via nitroxide‐mediated polymerization to obtain polymer with narrow polydispersity index (PDI) and controlled molecular weight. In this study, we tested the stem cell growth on glass coverslips coated with peptide (extracellular matrix related) conjugated‐PVBA‐St. Up on initial screening, three active peptides were identified suitable for pluripotent stem cells expansion and self‐renewal. The long‐term culture of three pluripotent stem cell lines (H7, HUES‐7 and DF699) further confirmed the applicability and robustness of the copolymer system.Support or Funding InformationFunding was provided by the Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore). VMR was supported by funding from National Institutes of Health grant (NIH/NIDDK RO1DK104791)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Environmental sustainability assessment of the manufacturing process of a biological active pharmaceutical ingredient

AbstractBACKGROUNDAlthough the biopharmaceutical market grows rapidly, the environmental impacts of biopharmaceutical manufacturing have been hardly evaluated. The aim of the study is to comprehensively assess the environmental sustainability of a biologically‐produced active pharmaceutical ingredient (API) manufacturing process. For this purpose, an exergy analysis and an (exergetic) life cycle assessment were performed using primary (foreground system) and modeled data (background system). Infliximab, a monoclonal antibody, was considered.RESULTSThe fermentation and its raw materials' supply chain was determined to have the highest environmental impact, since the culture media requires chemicals and complex compounds such as animal‐derived materials (ADMs). The total impact per 100 mg of API produced ranged between 18.6 and 101.8 MJex and 2.0–3.1 kg CO2‐eq, mainly as a function of the ADMs' source. A comparison of infliximab and ustekinumab, a biopharmaceutical produced without ADMs, showed that eliminating these materials can reduce the cumulative resource consumption of monoclonal antibody manufacturing by up to 7.5 times. The fermentation is the process taking the longest and the heating, ventilation and air conditioning system which support fermentation areas consumes approximately 75% of all plant electricity use.CONCLUSIONSCulture media nutrients and cleanroom requirements are driving factors in the environmental impact of biological API manufacturing. Obtaining more precise data of the bioprocesses behind these nutrients' supply chain is crucial to corroborating this study's results. © 2019 Society of Chemical Industry

Bioengineering the microanatomy of human skin.

Recreating the structure of human tissues in the laboratory is valuable for fundamental research, testing interventions, and reducing the use of animals. Critical to the use of such technology is the ability to produce tissue models that accurately reproduce the microanatomy of the native tissue. Current artificial cell‐based skin systems lack thorough characterisation, are not representative of human skin, and can show variation. In this study, we have developed a novel full thickness model of human skin comprised of epidermal and dermal compartments. Using an inert porous scaffold, we created a dermal construct using human fibroblasts that secrete their own extracellular matrix proteins, which avoids the use of animal‐derived materials. The dermal construct acts as a foundation upon which epidermal keratinocytes were seeded and differentiated into a stratified keratinised epithelium. In‐depth morphological analyses of the model demonstrated very close similarities with native human skin. Extensive immunostaining and electron microscopy analysis revealed ultrastructural details such as keratohyalin granules and lamellar bodies within the stratum granulosum, specialised junctional complexes, and the presence of a basal lamina. These features reflect the functional characteristics and barrier properties of the skin equivalent. Robustness and reproducibility of in vitro models are important attributes in experimental practice, and we demonstrate the consistency of the skin construct between different users. In summary, a new model of full thickness human skin has been developed that possesses microanatomical features reminiscent of native tissue. This skin model platform will be of significant interest to scientists researching the structure and function of human skin.

Preparation and investigation of novel SrCl2/DCMC-modified (via DOPA) decellularized arteries with excellent physicochemical properties and cytocompatibility for vascular scaffolds.

A new method of fabricating vascular scaffolds was designed in this article by crosslinking the porcine arteries using dialdehyde carboxymethyl (DCMC) and further introducing the Sr element on the surface of modified arteries using DOPA. DCMC had been selected as an ideal crosslinking reagent for its excellent cytobiocompatibility and suitable chemical reactivity. Unfortunately, the endothelialization of biological vascular scaffolds fixed by DCMC was unsatisfactory. To overcome this deficiency, the Sr element was introduced onto arteries to improve the endothelialization of fixed arteries due to the Sr element being able to promote the expression of vascular endothelial growth factor (VEGF) being crucial for growth and proliferation of HUVECs. After modifying and crosslinking, their chemical structures, mechanical properties, stability, and cytocompatibility were examined. Our findings demonstrated that DCMC could improve the mechanical properties of animal-derived materials successfully and possess suitable biocompatibility compared with glutaraldehyde (GA). The Sr element can easily be introduced onto the surface of DCMC modified arteries by DOPA. Compared with purely DCMC-crosslinked ones, SrCl2/DCMC modification has no significant effect on the mechanical strength of fixed arteries, but a slight tendancy to improve the stability of fixed samples in D-Hanks solution. MTT assay and fluorescence tests implied that SrCl2/DCMC modification could effectively stimulate HUVECs' adhesion and proliferation, and thus promote the endothelialization process of fixed arteries. SrCl2/DCMC-modified arteries with excellent physicochemical properties and appealing HUVEC-cytocompatibility should be promising materials for fabricating vascular scaffolds.