Animal Cell Culture Research Articles

Review in edible materials for sustainable cultured meat: scaffolds and microcarriers production

SummaryCultured meat, also known as laboratory‐grown or cell‐based meat, is a promising concept in the food industry that involves culturing animal cells in vitro to produce meat products, bypassing the need for traditional animal rearing and slaughter. This technology offers solutions to key challenges associated with conventional meat production, including environmental sustainability, animal welfare and food security. However, the cultured meat industry must overcome significant hurdles to achieve commercial viability. Scaling up production to meet the increasing demand for meat is a considerable challenge, requiring a scalable edible scaffold, eliminating the need for separate scaffold removal, and ensuring product safety. Various edible materials, including polysaccharides and proteins, along with fabrication techniques like electrospinning and 3D printing, are explored in this review article.

Challenging cultured meat naturalness perceptions: The role of consumers’ mindset

Cultured meat, produced through in vitro cultivation of animal cells, has emerged as a promising solution to environmental, health, and ethical issues resulting from conventional meat production. However, acceptance remains a crucial challenge, significantly influenced by perceptions of unnaturalness. Previous research has demonstrated the limited success of messaging strategies aimed at countering these perceptions. Across two experimental studies, this research breaks new ground by examining these strategies through the lens of mindset theory—i.e., beliefs about the fixedness or malleability of human attributes. In Study 1, we present findings illustrating that a strategy challenging the importance of naturalness is effective at increasing cultured meat acceptance among consumers with a growth mindset. In Study 2, we demonstrate how complementing such messaging strategy with a specific form of creative narrative can make it effective among consumers with a fixed mindset too. Our findings are informative theoretically, extending mindset and narrative theories to the context of cultured meat and, practically, examining the effectiveness of different communication strategies in driving consumer acceptance of the product.

FLASH Genome Editing Pipeline: An Efficient and High-Throughput Method to Construct Arrayed CRISPR Library for Plant Functional Genomics.

CRISPR/Cas9 genome editing is a revolutionary technology for plant functional genomics and crop breeding. In this system, the Cas9 nuclease is directed by a guide RNA (gRNA) to cut the DNA target and introduce mutation through error-prone DNA break repair. Owing to its simplicity, CRISPR/Cas9-mediated targeted gene knockout is widely used for high-throughput genetic screening in animal cell cultures and bacteria. However, high-throughput genetic screening using CRISPR/Cas9 is still challenging in plants. We recently established a new approach, named the FLASH genome editing pipeline, to construct an arrayed CRISPR library in plants. In this pipeline, a set of 12 PCR fragments with different lengths (referred to as FLASH tags) are used to index the Cas9/gRNA vectors. Subsequently, a mixture of 12 Agrobacterium strains, in which each strain contained a FLASH-tag indexed vector, was transformed into rice plants. As a result, a unique link between the target gene/gRNA and FLASH tag is generated, which allows reading gRNA information in bacterial strains and gene-edited plants using regular PCR and gel electrophoresis. This protocol includes step-by-step instructions for gRNA design, high throughput assembly of FLASH-tag indexed Cas9/gRNA plasmids, Agrobacterium-mediated transformation of 12 indexed plasmids, and fast assignment of target gene information in primary transformants. The arrayed CRISPR library described here is suitable for small- to large-scale genetic screening and allows fast and comprehensive gene function discovery in plants. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Assembly of FLASH-tag-indexed Cas9/gRNA plasmids Basic Protocol 2: Preparation of the Cas9/gRNA plasmid library Basic Protocol 3: Library preparation of Agrobacterium strains and mixing FLASH-tag indexed strains Basic Protocol 4: Grouped transformation and assignments of gRNA information of gene-edited plants.

Psychobiotics: Are they the future intervention for managing depression and anxiety? A literature review.

Mental health is a public health concern among professional organizations, clinicians, and consumers alike, especially in light of the COVID-19 pandemic. Indeed, the World Health Organization has identified mental health as an epidemic of the 21st century contributing to the global health burden, which highlights the urgency to develop economical, accessible, minimally invasive interventions to effectively manage depression, anxiety, and stress. Nutritional approaches, including the use of probiotics and psychobiotics to manage depression and anxiety, have elicited interest in recent years. This review aimed to summarize evidence from studies including animal models, cell cultures, and human subjects. Overall, the current evidence suggests that 1) Specific strains of probiotics can reduce depressive symptoms and anxiety; 2) Symptoms may be reduced through one or more possible mechanisms of action, including impact on the synthesis of neurotransmitters such as serotonin and GABA, modulation of inflammatory cytokines, or enhancing stress responses through effects on stress hormones and the HPA axis; and 3) While psychobiotics may offer therapeutic benefits to manage depression and anxiety, further research, particularly human studies, is needed to better characterize their mode of action and understand optimal dosing in the context of nutritional interventions.

Efficiency Comparative Approach of Plant-Produced Monoclonal Antibodies against Rabies Virus Infection.

Rabies encephalitis is a fatal zoonotic viral disease caused by the neurotropic rabies virus. It remains a major public health concern as it causes almost 100% fatality and has no effective medication after the onset of the disease. However, this illness is preventable with the timely administration of effective post-exposure prophylaxis (PEP) consisting of the rabies vaccine and passive immune globulins (HRIG and ERIG). Recently, conventional PEP has been shown to have many limitations, resulting in little support for these expensive and heterologous globulins. Monoclonal antibody (mAb) production via recombinant technology in animal and human cell cultures, as well as a plant-based platform, was introduced to overcome the costly and high-tech constraints of former preparations. We used transient expression technology to produce two mAbs against the rabies virus in Nicotiana benthamiana and compared their viral neutralizing activity in vitro and in vivo. The expression levels of selective mAbs E559 and 62-71-3 in plants were estimated to be 17.3 mg/kg and 28.6 mg/kg in fresh weight, respectively. The plant-produced mAbs effectively neutralized the challenge virus CVS-11 strain in a cell-based RFFIT. In addition, the combination of these two mAbs in a cocktail protected hamsters from rabies virus infection more effectively than standard HRIG and ERIG. This study suggests that the plant-produced rabies antibody cocktail has promising potential as an alternative biological to polyclonal RIG in rabies PEP.

Product safety aspects of plant molecular farming.

Plant molecular farming (PMF) has been promoted since the 1990s as a rapid, cost-effective and (most of all) safe alternative to the cultivation of bacteria or animal cells for the production of biopharmaceutical proteins. Numerous plant species have been investigated for the production of a broad range of protein-based drug candidates. The inherent safety of these products is frequently highlighted as an advantage of PMF because plant viruses do not replicate in humans and vice versa. However, a more nuanced analysis of this principle is required when considering other pathogens because toxic compounds pose a risk even in the absence of replication. Similarly, it is necessary to assess the risks associated with the host system (e.g., the presence of toxic secondary metabolites) and the production approach (e.g., transient expression based on bacterial infiltration substantially increases the endotoxin load). This review considers the most relevant host systems in terms of their toxicity profile, including the presence of secondary metabolites, and the risks arising from the persistence of these substances after downstream processing and product purification. Similarly, we discuss a range of plant pathogens and disease vectors that can influence product safety, for example, due to the release of toxins. The ability of downstream unit operations to remove contaminants and process-related toxic impurities such as endotoxins is also addressed. This overview of plant-based production, focusing on product safety aspects, provides recommendations that will allow stakeholders to choose the most appropriate strategies for process development.

Establishment and characterization of a novel kidney cell line derived from the common bottlenose dolphin.

Common bottlenose dolphin (Tursiops truncatus) is a well-known cetacean species that inhabits temperate and tropical seas worldwide. Limited supply and poor quality of samples hinder the investigation of the effects of various pathogens and environmental pollutants on this cetacean species. Cultured cells are useful for experimental studies; however, no cell lines derived from cetaceans are generally available. Therefore, in this study, we established a novel kidney cell line, TK-ST, derived from T. truncatus. Primary cells exhibited the morphological characteristics of epithelial and fibroblast cells, but their immortalization and passaging resulted in a predominantly epithelial cell morphology. TK-ST was immortalized using the large T SV40 antigen and human telomerase reverse transcriptase and exhibited long-term stable cell growth. TK-ST cells are generally cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum at 37°C and 5% CO2 but can also be cultured in 5-20% fetal bovine serum and several other classical media commonly used for common animal cell culture. TK-ST cells were found to be susceptible to several viruses, including the dolphin morbillivirus (most important virus in cetaceans), and exhibited cytopathic effects, facilitating the replication of the dolphin morbillivirus. Furthermore, mRNA expression levels of cytokine genes were increased in TK-ST cells after stimulation with lipopolysaccharides and poly(I:C). Therefore, the novel TK-ST cell line derived in this study can potentially be used for further in vitro studies on cetaceans.

Pyrrolizidine plant extracts as diagnostic biochemical markers on therapeutic consequences

The consumption of cereals and bakery products contaminated with seeds of species containing Pyrrolizidine alkaloids (PAs) has been involved in mass poisonings in rural areas of Afghanistan, India, South Africa and the former USSR. Poisoning can manifest as acute or subacute veno-occlusive disease with specific symptoms such as persistent hepatomegaly, leading to cirrhosis or shown to have genotoxic, mutagenic, teratogenic and carcinogenic effects. Several such toxicity testing of PAs have been done in cultured animal cells with reference to mice model, to better understand the pathway of metabolic impairement. Also the pluripotent proprty of mice bone marrow cells show better analyzing capacity of differentiated cell lineages. Therefore this pre-analytic review aims to focus the risk evaluation methods of PA exposure on current knowledge and recent advances of metabolic pathways of different PAs. In addition, this review provides perspectives of precision toxicity assessment strategies and biomarker development for the risk control of human intoxication by PAs with eventual categorization of pyrrole-protein adducts like serum Alanine Transferase (ALT), Glutathione transferase (GSH-T), Alkaline Phosphatase (ALP) in different cell lineages grown by lab-based cell culture techniques.

Low CO2 partial pressure steers CHO cells into a defective metabolic state.

The accumulation of carbon dioxide during large-scale culture of animal cells brings adverse effects, appropriate aeration strategies alleviate CO2 accumulation while improper reactor operation may lead to the presence of low CO2 partial pressure (pCO2) condition as occurs in many industrial cases. Thus, this study aims to reveal the in-depth influence of low pCO2 on Chinese Hamster Ovary (CHO) cells for providing a reference for design space determination of CO2 control with regard to the Quality by Design (QbD) guidelines. The headspace air over purging caused the ultra-low pCO2 (ULC) where the monoclonal antibody production as well as the aerobic metabolic activity were reduced. Intracellular metabolomics analysis indicated a less efficient aerobic glucose metabolic state under ULC conditions. Based on the increase of intracellular pH and lactate dehydrogenase activity, the shortage of intracellular pyruvate could be the cause of the deficient aerobic metabolism, which could be partially mitigated by pyruvate addition under ULC conditions. Finally, a semi-empirical mathematical model was used to better understand, predict and control the occurrence of extreme pCO2 conditions during the cultures of CHO cells. Low pCO2 steers CHO cells into a defective metabolic state. A predictive relation among pCO2, lactate, and pH control was applied to get new insights into CHO cell culture for better and more robust metabolic behavior and process performance and the determination of QbD design space for CO2 control.

On the nature of picobirnaviruses.

The picobirnaviruses (Picobirnaviridae, Picobirnavirus, PBVs) are currently thought to be animal viruses, as they are usually found in animal stool samples. However, no animal model or cell culture for their propagation has yet been found. In 2018, a hypothetical assumption about PBVs belonging to prokaryotic viruses was put forward and experimentally substantiated. This hypothesis is based on the presence of Shine-Dalgarno sequences in the genome of all PBVs before three reading frames (ORF) at the ribosomal binding site, with which the prokaryotic genome is saturated, while in the eukaryotic genome such regions occur with low frequency. The genome saturation with the Shine-Dalgarno sequences, as well as the preservation of this saturation in the progeny, according to scientists, allows us to attribute PBVs to prokaryotic viruses. On the other hand, there is a possibility that PBVs belong to viruses of eukaryotic hosts - fungi or invertebrates, since PBV-like sequences similar to the genome of fungal viruses from the families of mitoviruses and partitiviruses have been identified. In this regard, the idea arose that, in terms of reproduction mode, PBVs resemble fungal viruses. The divergence of views on the true PBV host(s) has sparked discussions among scientists and required further research to elucidate their nature. The review highlights the results of the search for a PBV host. The reasons for the occurrence of atypical sequences among the PBV genome sequences that use an alternative mitochondrial code of lower eukaryotes (fungi and invertebrates) for the translation of viral RNA-dependent RNA polymerase (RdRp) instead of the standard genetic code are analyzed. The purpose of the review was to collect arguments in support of the hypothesis about the phage nature of PBVs and to find the most realistic explanation of the reasons for identifying non-standard genomic sequences for PBVs. Based on the hypothesis about the genealogical relationship of PBVs with RNA viruses from other families with similar segmented genomes, such as Reoviridae, Cystoviridae, Totiviridae and Partitiviridae, virologists support the assumption of a decisive role in the origin of atypical PBV-like reassortment strains between PBVs and viruses of the listed families. The collected arguments given in this review indicate a high probability of a phage nature of PBVs. The data presented in the review show that the belonging of PBV-like progeny to prokaryotic or eukaryotic viruses is determined not only by its genome saturation level with a prokaryotic motif, standard or mitochondrial genetic code. The primary structure of the gene encoding the viral capsid protein responsible for the presence or absence of specific proteolytic properties of the virus that determine its ability for independent horizontal transmission into new cells may also be a decisive factor.

Fulfilling the Promises of Fermentation-Derived Foods

Fermentation-derived foods (FDFs) such as cultured animal cells, edible microbial biomass, and recombinant food proteins have recently seen a notable surge in both investment and public interest. FDFs have been promoted as a more ethical, resilient, and environmentally sustainable food alternative to animal-derived foods such as meat, eggs, and dairy. However, some of the manufacturing process choices and sociopolitical assumptions made by actors in this space could risk undermining the goals of the technology as stated above. This article highlights five aspects of FDFs that should be carefully considered if they are to assist human society successfully transition to a more ethical, sustainable, and resilient food production system: (1) sustainable nitrogen fixation and management, (2) transitioning away from sugar feedstocks, (3) realistic expectations of consumer adoption rate, (4) careful consideration of the role of intellectual property, and (5) greater emphasis on food products that do not require cold or frozen storage.

Exploring Toxoplasma gondii´s Biology within the Intestinal Epithelium: intestinal-derived models to unravel sexual differentiation.

A variety of intestinal-derived culture systems have been developed to mimic in vivo cell behavior and organization, incorporating different tissue and microenvironmental elements. Great insight into the biology of the causative agent of toxoplasmosis, Toxoplasma gondii, has been attained by using diverse in vitro cellular models. Nonetheless, there are still processes key to its transmission and persistence which remain to be elucidated, such as the mechanisms underlying its systemic dissemination and sexual differentiation both of which occur at the intestinal level. Because this event occurs in a complex and specific cellular environment (the intestine upon ingestion of infective forms, and the feline intestine, respectively), traditional reductionist in vitro cellular models fail to recreate conditions resembling in vivo physiology. The development of new biomaterials and the advances in cell culture knowledge have opened the door to a next generation of more physiologically relevant cellular models. Among them, organoids have become a valuable tool for unmasking the underlying mechanism involved in T. gondii sexual differentiation. Murine-derived intestinal organoids mimicking the biochemistry of the feline intestine have allowed the generation of pre-sexual and sexual stages of T. gondii for the first time in vitro, opening a window of opportunity to tackling these stages by "felinizing" a wide variety of animal cell cultures. Here, we reviewed intestinal in vitro and ex vivo models and discussed their strengths and limitations in the context of a quest for faithful models to in vitro emulate the biology of the enteric stages of T. gondii.

Pathophysiology of Retinopathy of Prematurity.

Retinopathy of prematurity (ROP) is a complex disease involving development of the neural retina, ocular circulations, and other organ systems of the premature infant. The external stresses of the ex utero environment also influence the pathophysiology of ROP through interactions among retinal neural, vascular, and glial cells. There is variability among individual infants and presentations of the disease throughout the world, making ROP challenging to study. The methods used include representative animal models, cell culture, and clinical studies. This article describes the impact of maternal-fetal interactions; stresses that the preterm infant experiences; and biologic pathways of interest, including growth factor effects and cell-cell interactions, on the complex pathophysiology of ROP phenotypes in developed and emerging countries.

L-Lactate treatment by photosynthetic cyanobacteria expressing heterogeneous l-lactate dehydrogenase

l-Lactate is a major waste compound in cultured animal cells. To develop a sustainable animal cell culture system, we aimed to study the consumption of l-lactate using a photosynthetic microorganism. As genes involved in l-lactate utilization were not found in most cyanobacteria and microalgae, we introduced the NAD-independent l-lactate dehydrogenase gene from Escherichia coli (lldD) into Synechococcus sp. PCC 7002. The lldD-expressing strain consumed l-lactate added to basal medium. This consumption was accelerated by expression of a lactate permease gene from E. coli (lldP) and an increase in culture temperature. Intracellular levels of acetyl-CoA, citrate, 2-oxoglutarate, succinate, and malate, and extracellular levels of 2-oxoglutarate, succinate, and malate, increased during l-lactate utilization, suggesting that the metabolic flux from l-lactate was distributed toward the tricarboxylic acid cycle. This study provides a perspective on l-lactate treatment by photosynthetic microorganisms, which would increase the feasibility of animal cell culture industries.

A COVID-19 vaccine candidate based on SARS-CoV-2 spike protein and immune-stimulating complexes

Spike protein from SARS-CoV-2, the etiologic agent of the COVID-19 pandemic disease, constitutes a structural protein that proved to be the main responsible for neutralizing antibody production. Thus, its sequence is highly considered for the design of candidate vaccines. Animal cell culture represents the best option for the production of subunit vaccines based on recombinant proteins since they introduce post-translational modifications that are important to mimic the natural antigenic epitopes. Particularly, the human cell line HEK293T has been explored and used for the production of biotherapeutics since the products derived from them present human-like post-translational modifications that are important for the protein's activity and immunogenicity. The aim of this study was to produce and characterize a potential vaccine for COVID-19 based on the spike ectodomain (S-ED) of SARS-CoV-2 and two different adjuvants: aluminum hydroxide (AH) and immune-stimulating complexes (ISCOMs). The S-ED was produced in sHEK293T cells using a 1-L stirred tank bioreactor operated in perfusion mode and purified. S-ED characterization revealed the expected size and morphology. High N-glycan content was confirmed. S-ED-specific binding with the hACE2 (human angiotensin-converting enzyme 2) receptor was verified. The immunogenicity of S-ED was evaluated using AH and ISCOMs. Both formulations demonstrated the presence of anti-RBD antibodies in the plasma of immunized mice, being significantly higher for the latter adjuvant. Also, higher levels of IFN-γ and IL-4 were detected after the ex vivo immune stimulation of spleen-derived MNCs from ISCOMs immunized mice. Further analysis confirmed that S-ED/ISCOMs elicit neutralizing antibodies against SARS-CoV-2. KEY POINTS: Trimeric SARS-CoV-2 S-ED was produced in stable recombinant sHEK cells in serum-free medium. A novel S-ED vaccine formulation induced potent humoral and cellular immunity. S-ED formulated with ISCOMs adjuvant elicited a highly neutralizing antibody titer.

Are encapsulated pesticides less harmful to human health than their conventional alternatives? Protocol for a systematic review of in vitro and in vivo animal model studies

BackgroundThe gradual increase in the global population has led to the rising demand for agricultural products worldwide. This required the introduction of environment- and public health-friendly advanced technologies for plant protection to guard yields from pest destruction in a sustainable way. Encapsulation technology is a promising procedure to increase the effectiveness of pesticide active ingredients while reducing human exposure and environmental impact. Despite the presumed favorable properties of encapsulated pesticide formulations on human health, it is necessary to systematically assess whether they are less harmful to human health than conventional pesticide products. ObjectivesWe aim to systematically review the literature to answer the question of whether micro- or nano-encapsulated pesticide formulations exert different degrees of toxicity than their conventional (not-encapsulated) counterparts in in vivo animal and in vitro (human, animal, and bacterial cell) non-target models. The answer is important to estimate the possible differences in the toxicological hazards of the two different types of pesticide formulations. Because our extracted data will come from different models, we also aim to perform subgroup analyses to investigate how toxicity varies across different models. A pooled toxicity effect estimate will also be performed by meta-analysis when appropriate. MethodsThe systematic review will follow the guidelines developed by the National Toxicology Program’s Office of Health Assessment and Translation (NTP/OHAT). The protocol adheres to the Preferred Reporting Items for Systematic Reviews and meta-analyses Protocol (PRISMA-P) statement. PubMed (NLM), Scopus (Elsevier), Web of Science Core Collection (Clarivate), Embase (Elsevier), and Agricola (EBSCOhost) electronic databases will be comprehensively searched in September 2022 to identify eligible studies using multiple search terms of “pesticide”, “encapsulation” and “toxicity” along with their synonyms and other words that are semantically related. The reference lists of all eligible articles and retrieved reviews will be manually screened to identify additional relevant papers. Eligibility criteriaWe will include peer-reviewed experimental (non-target in vivo animal model and in vitro human, animal, and bacterial cell cultures) studies published as full-text articles in English language that simultaneously investigate the effect of any micro- or nano-encapsulated pesticide formulation, applied in all ranges of concentrations, duration, and routes of exposure, and its corresponding active ingredient(s) or its conventional non-encapsulated product formulation(s) used in the same ranges of concentrations, duration, and routes of exposure on the same pathophysiological outcome. We will exclude studies that examine pesticidal activity on target organisms, cultures of cells isolated from target organisms exposed in vivo or in vitro, and those using biological materials isolated from target organisms/cells. Study appraisal and synthesisStudies identified by the search will be screened and managed according to the review inclusion and exclusion criteria in the Covidence systematic review tool by two reviewers, who will also blindly extract the data and assess the risk of bias of included studies. The OHAT risk of bias tool will be applied to evaluate the quality and risk of bias in the included studies. Study findings will be synthesized narratively by important features of the study populations, design, exposure, and endpoints. If findings make it possible, a meta-analysis will be performed on identified toxicity outcomes. To rate the certainty in the body of evidence, we will use the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach.

Anticancer activity of lycopene in HT-29 colon cancer cell line

An inverse association between serum lycopene levels and the risk of cancers has been pointed out by many prospective and retrospective epidemiological studies which prompted more studies to be performed on animal models and cell cultures in order to test this hypothesis. The aim of the present study was to evaluate the antiproliferative and pro-apoptotic effect of lycopene on colon cancer HT-29 cell line. The effect of lycopene on the viability of HT-29 cell line was investigated using XTT assay. The levels of Bcl-2, cleaved caspase 3, BAX, cleaved PARP, and 8-oxo-dG in lycopene-treated HT-29 cells were measured using ELISA. Gamma-H2AX and cytochrome c expression was assessed semi-quantitatively using immunofluorescence staining. Lycopene at doses of 10 and 20 μM produced a significant antiproliferative effect on HT-29 cells compared to the control (p < 0.05). The IC50 value of lycopene in HT-29 cells was found to be 7.89 μM for 24 h. Lycopene (7.89 μM) significantly elevated cleaved caspase 3 (p <0.01), BAX, and cleaved PARP, 8-oxo-dG levels (p<0.05). The levels of γ-H2AX foci are significantly higher while the levels of cytochrome-c are lower (p<0.05) in lycopene-treated HT-29 cells. These results indicate that lycopene has an antiproliferative apoptotic and genotoxic effect on HT-29 colon cancer cell line.

One-Pot Radiosynthesis of [18F]Anle138b—5-(3-Bromophenyl)-3-(6-[18F]fluorobenzo[d][1,3]dioxol-5-yl)-1H-pyrazole—A Potential PET Radiotracer Targeting α-Synuclein Aggregates

Availability of PET imaging radiotracers targeting α-synuclein aggregates is important for early diagnosis of Parkinson's disease and related α-synucleinopathies, as well as for the development of new therapeutics. Derived from a pyrazole backbone, 11C-labelled derivatives of anle138b (3-(1,3-benzodioxol-5-yl)-5-(3-bromophenyl)-1H-pyrazole)-an inhibitor of α-synuclein and prion protein oligomerization-are currently in active development as the candidates for PET imaging α-syn aggregates. This work outlines the synthesis of a radiotracer based on the original structure of anle138b, labelled with fluorine-18 isotope, eminently suitable for PET imaging due to half-life and decay energy characteristics (97% β+ decay, 109.7 min half-life, and 635 keV positron energy). A three-step radiosynthesis was developed starting from 6-[18F]fluoropiperonal (6-[18F]FP) that was prepared using (piperonyl)(phenyl)iodonium bromide as a labelling precursor. The obtained 6-[18F]FP was used directly in the condensation reaction with tosylhydrazide followed by 1,3-cycloaddition of the intermediate with 3'-bromophenylacetylene eliminating any midway without any intermediate purifications. This one-pot approach allowed the complete synthesis of [18F]anle138b within 105 min with RCY of 15 ± 3% (n = 3) and Am in the range of 32-78 GBq/µmol. The [18F]fluoride processing and synthesis were performed in a custom-built semi-automated module, but the method can be implemented in all the modern automated platforms. While there is definitely space for further optimization, the procedure developed is well suited for preclinical studies of this novel radiotracer in animal models and/or cell cultures.

Review on cultured meat: ethical alternative to animal industrial farming

It is estimated that around 8% of the entire world population identifies themselves as vegetarians. That leaves us with statistics of more than 90% of the world's population prefers a meat-based diet. Now with the alarmingly increasing population of over 7.8 billion, the demand for meat is skyrocketing. Due to the high demand, factors like land, water and greenhouse emission are becoming unsustainable. Even after taking all these concerns into account, why is it so difficult for us to eliminate/reduce meat from our diet? How is the future generation going to satisfy their meat cravings? However, through recent research, it seems there is an alternative, a solution to all these concerns, cultured/ lab-grown meat. Cultured meat is meat produced by in vitro cell culture of animal cells, instead of from slaughtered animals using various tissue engineering techniques. This invitro meat technology is still at its elementary stages but shows great potential in curbing the problem of livestock overcrowding and its impact on the environment. Cultured meat seems to promise a sustainable and reliable alternative for its consumers. This review aimed to provide an elaborate comprehension of the entire process of the production, processing and commercialization of cultured meat, and also provides an insight on the possible obstacles it encounters, especially overcoming ethical conundrums and social prejudice.

PGEX2-T based β defensin primer designing and isolation of neutrophil, macrophage, bone marrow and mesenchymal stem cells

Defensins are antimicrobial peptides that act mainly by disrupting the structure of bacterial cell membranes and are found in many compartments of the body. Defensins play a central role in defense against pathogens and they are considered part of the innate immune response. Material and methods-Reagents: Animal cell culture media - Growth medium D-MEM, RPMI and alpha -MEM was purchased from gibco, CA, USA. Fetal bovine serum (FBS)was obtained from Invitrogen, CA, USA. DAPI, agarose was purchased from SRL, India. Penicillium streptomycin solution was obtained from himedia, Mumbai, India. Animal modal: In bred populations of Swiss albino mice of either sex and at 6-8 weeks of age were used. Mice obtained from the Animal House, All India Institute of Medical Science, NewDelhi-110029 and India Standardization, Isolation and purification and characterization of bone marrow from Swiss albino mice. Bone marrow is consisting of multiple type of stem cells such as -Mesenchymal stem cells (MSCs) which are multipotent stem cells that have the potential to self-renew and differentiate into a variety of specialized cell types such as osteoblasts, chondrocytes, adipocytes, and neurons. Result: Cells were harvested by adherence purification, the adherence of cells which is a symbol of macrophage and acentric nucleus is visible in DAPI stain. The extended cytoplasmic surface and CD14 positive staining show the purified macrophage as 98% purity. BM-MSC were harvested by adherence purification, the adherence of cells to the plastic culture dish surface is a symbol of Mesenchymal stem cell and nucleusis visible in DAPI stain. Conclusion: This procedure is widely used to study macrophage biology since moderate numbers of resident, unstimulated macrophages can easily be obtained fromthe peritonealcavity.