Emerging noninvasive methylation biomarkers of cancer prognosis and drug response prediction.

Cancer is the second leading cause of death worldwide being responsible for 9.6 million deaths in 2018. Epigenetic alterations are key in directing the aberrant expression of tumor-associated genes that drive cellular malignant transformation and cancer progression. Among epigenetic alterations, DNA methylation is the most deeply studied one in relation to environmental exposure. Tissue biopsies have traditionally been the main procedure by which a small sample of body tissue is excised to confirm cancer diagnosis or to indicate the primary site when cancer has spread. In contrast, the analysis of circulating tumor-derived material, or tumor circulome, by means of liquid biopsy of peripheral blood, urine, saliva or sputum is a noninvasive, fast and reproducible alternative to tissue biopsy. Recently, the assessment of epigenetic alterations such as DNA methylation and hydroxymethylation in circulating free DNA has been proved possible. These marks can be associated to prognosis and response to a variety of treatments including chemotherapy, hormonotherapy or immunotherapy. Epigenetic biomarkers may offer some advantages over RNA or genetic biomarkers given their stability in bodily fluids and their high tissue-specificity. While many challenges are still ahead, the unique advantages of these types of biomarkers is urging the scientific community to persevere in their clinical validation and integration into reliable prediction models. This review aims at recapitulating the emerging noninvasive DNA methylated biomarkers of importance for prediction of prognosis and drug response in cancer.

Preclinical models in head and neck squamous cell carcinoma.

Head and neck cancer is the sixth most frequent cancer type. Drug resistance and toxicity are common challenges of the existing therapies, making the development of reliable preclinical models essential for the study of the involved molecular mechanisms as well as for eventual intervention approaches that improve the clinical outcome. Preclinical models of head and neck squamous cell carcinoma have been traditionally based on cell lines and murine models. In this review, we will go over the most frequently used preclinical models, from immortalised-cell and primary tumour cultures in monolayer or 3D, to the currently available animal models. We will scrutinise their efficiency in mimicking the molecular and cellular complexity of head and neck squamous cell carcinoma. Finally, the challenges and the opportunities of other envisaged putative approaches, as well as the potential of the preclinical models to further develop personalised therapies will be discussed.

High IGKC-Expressing Intratumoral Plasma Cells Predict Response to Immune Checkpoint Blockade.

Resistance to Immune Checkpoint Blockade (ICB) constitutes the current limiting factor for the optimal implementation of this novel therapy, which otherwise demonstrates durable responses with acceptable toxicity scores. This limitation is exacerbated by a lack of robust biomarkers. In this study, we have dissected the basal TME composition at the gene expression and cellular levels that predict response to Nivolumab and prognosis. BCR, TCR and HLA profiling were employed for further characterization of the molecular variables associated with response. The findings were validated using a single-cell RNA-seq data of metastatic melanoma patients treated with ICB, and by multispectral immunofluorescence. Finally, machine learning was employed to construct a prediction algorithm that was validated across eight metastatic melanoma cohorts treated with ICB. Using this strategy, we have unmasked a major role played by basal intratumoral Plasma cells expressing high levels of IGKC in efficacy. IGKC, differentially expressed in good responders, was also identified within the Top response-related BCR clonotypes, together with IGK variants. These results were validated at gene, cellular and protein levels; CD138+ Plasma-like and Plasma cells were more abundant in good responders and correlated with the same RNA-seq-defined fraction. Finally, we generated a 15-gene prediction model that outperformed the current reference score in eight ICB-treated metastatic melanoma cohorts. The evidenced major contribution of basal intratumoral IGKC and Plasma cells in good response and outcome in ICB in metastatic melanoma is a groundbreaking finding in the field beyond the role of T lymphocytes.

Physiological and biochemical responses driven by different UV-visible radiation in Osmundea pinnatifida (Hudson) Stackhouse (Rhodophyta).

Light, or visible radiation, serves as a source of energy for photosynthesis of plants and most algae. In addition, light and ultraviolet radiation (UV-A and UV-B) act as a biological signal, triggering several cellular processes that are mediated by photoreceptors. The aim of this study was to evaluate the physiological and biochemical responses of Osmundea pinnatifida driven by different radiations through putative photoreceptors. For this, O. pinnatifida was grown under different radiation treatments composed by high intensity of light emitted by a low pressure sodium lamp (SOX), aiming to saturate photosynthesis, which was supplemented by low intensities of visible (red, green and blue) and ultraviolet radiation (UV-A and UV-B), in order to activate photoreceptors. Growth rates, photosynthesis, antioxidant activity, polyphenols, soluble proteins, phycobiliproteins, mycosporine-like amino acids (MAAs) and carotenoids were evaluated during the experiment. Complementary UV-A radiation positively influenced growth rates after 15 days of experiment, although the presence of a peak of blue light in this treatment can also have contributed. UV-B radiation increased the concentration of zeaxanthin and chlorophyll a. The blue light caused the accumulation of chlorophyll a, violaxanthin, phycoerythrin and polyphenols on different days of the experiment. Phycoerythrin also increased under green and red light conditions. Our results showed that some compounds can be modulated by different radiation, and the involvement of photoreceptors is suggested. In red algae, photoreceptors sensitive to red, green and blue light have been identified, however little is known about UV photoreceptors. The presence of photoreceptors sensitive to UV radiation in O. pinnatifida is discussed.

Loss of Canonical Notch Signaling Affects Multiple Steps in NK Cell Development in Mice.

Within the hematopoietic system, the Notch pathway is critical for promoting thymic T cell development and suppressing the B and myeloid lineage fates; however, its impact on NK lymphopoiesis is less understood. To study the role of Notch during NK cell development in vivo, we investigated different NK cell compartments and function in Rbp-Jkfl/flVav-Cretg/+ mice, in which Rbp-Jk, the major transcriptional effector of canonical Notch signaling, was specifically deleted in all hematopoietic cells. Peripheral conventional cytotoxic NK cells in Rbp-Jk-deleted mice were significantly reduced and had an activated phenotype. Furthermore, the pool of early NK cell progenitors in the bone marrow was decreased, whereas immature NK cells were increased, leading to a block in NK cell maturation. These changes were cell intrinsic as the hematopoietic chimeras generated after transplantation of Rbp-Jk-deficient bone marrow cells had the same NK cell phenotype as the Rbp-Jk-deleted donor mice, whereas the wild-type competitors did not. The expression of several crucial NK cell regulatory pathways was significantly altered after Rbp-Jk deletion. Together, these results demonstrate the involvement of canonical Notch signaling in regulation of multiple stages of NK cell development.

Emergence of NK-cell progenitors and functionally competent NK-cell lineage subsets in the early mouse embryo.

The earliest stages of natural killer (NK)-cell development are not well characterized. In this study, we investigated in different fetal hematopoietic tissues how NK-cell progenitors and their mature NK-cell progeny emerge and expand during fetal development. Here we demonstrate, for the first time, that the counterpart of adult BM Lin(-)CD122(+)NK1.1(-)DX5(-) NK-cell progenitor (NKP) emerges in the fetal liver at E13.5. After NKP expansion, immature NK cells emerge at E14.5 in the liver and E15.5 in the spleen. Thymic NK cells arise at E15.5, whereas functionally competent cytotoxic NK cells were present in the liver and spleen at E16.5 and E17.5, respectively. Fetal NKPs failed to produce B and myeloid cells but sustained combined NK- and T-lineage potential at the single-cell level. NKPs were also found in the fetal blood, spleen, and thymus. These findings show the emergence and expansion of bipotent NK/T-cell progenitor during fetal and adult lymphopoiesis, further supporting that NK/T-lineage restriction is taking place prethymically. Uncovering the earliest NK-cell developmental stages will provide important clues, helping to understand the origin of diverse NK-cell subsets, their progenitors, and key regulators.

Antibiotic residues and microbial contamination in pasteurized whole milk intended for human consumption.

Background and Aim: Milk contamination for human consumption is one of the biggest concerns worldwide. To prevent milk contamination, it is important to implement sustainable production practices that ensure animal health and guarantee veterinary drugs have been used properly. This study aimed to detect antibiotic residues and microbial contamination in commercially available pasteurized whole milk intended for human consumption. Materials and Methods: We conducted a cross-sectional study on all brands of pasteurized milk (n = 17) for human consumption in Medellín, Colombia, from February 30 to April 30, 2022. Six milk samples of each brand were collected every 15 days, resulting in 102 samples. IDEXX SNAPduo™ ST Plus test (IDEXX Laboratories Inc, Maine, USA) was used to detect cephalosporins residues to detect beta-lactam and tetracyclines. We detected mesophilic aerobic bacteria and coliforms using Chromocult Coliform Agar® (Merck KGaA, Darmstadt, Germany) and Plate-Count Agar® (Merck KGaA), respectively. Results: Beta-lactam residues were found in 24.4% of the brands. No tetracyclines or cephalosporins were detected. Mesophilic aerobic bacteria and coliform contamination were detected in 42.6% and 12.8% of the brands, respectively. No fecal coliform contamination was detected. Conclusion: This study demonstrated the presence of antibiotic residues and microbial contamination in commercially available pasteurized whole milk intended for human consumption in the study area, highlighting its potential public health implications.

Particleboard panels made with sugarcane bagasse waste-an exploratory study.

The reuse of natural fibers, in order to manufacture a new product, is already becoming popular due to the generation of a series of advantages in social areas. Sugarcane bagasse is a set of tangled fibers of cellulose, produced in large quantities due to increased acreage and industrialization of sugarcane resulting from public and private investments in production aimed for the alcohol industry. The aim of this study was to evaluate the feasibility of producing sheet timber manufacture from the sugarcane bagasse, analyzing mechanical strength properties. A form of metal sheet for the molding of 12 specimens based on sugarcane bagasse and industrialized resin was made. Soon after molding, specimens were submitted to a three-point bending test, with the aid of a press. The analysis of the results allowed to conclude that the tensile strength and the modulus of elasticity did not obtain the minimum values recommended by the standard. The tensile strength must be improved to allow panels to be useful for ordinary strength applications.

Immunological mechanisms underlying delayed-type hypersensitivity reactions to glatiramer acetate.

BACKGROUND: Delayed-type hypersensitivity to glatiramer acetate is rare, and the underlying immunological mechanisms are not completely understood. OBJECTIVE: To study the immunologic response in 2 patients with multiple sclerosis who developed maculopapular exanthema related with the administration of glatiramer acetate. METHODS: The allergologic study included general blood tests, viral serologic tests, and skin tests (patch and intradermal tests). The immunologic study was performed in skin biopsy specimens by immunohistochemistry and in the peripheral blood by flow cytometry and the lymphocyte transformation test. RESULTS: Skin test results were negative in both patients, and the diagnosis was confirmed by a drug provocation test. The evaluation of the acute phase showed an increase in the percentage of CD8 T lymphocytes (>50%) and the percentage of cells expressing skin-homing receptor (cutaneous lymphocyte-associated antigen) (>70%) and chemokine receptors (CCR4 and CXCR3) at T1. A positive proliferative response was observed in T lymphocytes (stimulation index [SI] = 3.5 in patient 1 and 3.59 in patient 2), especially the CD8(+) subpopulation (SI = 5.5 and 4.6 in patients 1 and 2, respectively), and NK lymphocytes (SI = 3.9 and 8.5 in patients 1 and 2, respectively) after glatiramer acetate stimulation. CONCLUSION: This study demonstrates the important role of T(H)1 cells expressing skin-homing receptors in delayed-type hypersensitivity reactions to glatiramer acetate. A lymphocyte transformation test revealed a specific glatiramer acetate recognition by T lymphocytes and NK lymphocytes.

Storage Duration Affects the Quantification of Oxidative Stress Markers in the Gastrocnemius, Heart, and Brain of Mice Submitted to a Maximum Exercise.

This study investigated the effect of sample storage duration on the quantification of oxidative stress markers in the gastrocnemius, heart, and brain of mice submitted to a maximum swimming exercise. Thiobarbituric acid reactive substances (TBARSs), protein carbonyl derivatives, total antioxidant capacity (TAC), and the activity of superoxide dismutase (SOD) and catalase (CAT) were quantified in fresh tissues and in samples stored at -80°C for 1, 3, or 6 months, from exercised (n = 13) and nonexercised mice (n = 13). Except for protein carbonyl derivatives in the heart, the exercise resulted in the modification of all markers in all fresh-evaluated samples (p < 0.001). The storage duration did not modify the effect of exercise on protein carbonyl derivatives and TAC. TBARS was stable for 3 months in the gastrocnemius and for 1 month in frozen heart and brain. Accordingly, the exercise effect on TBARS levels observed in fresh samples was absent in the gastrocnemius frozen for 6 months (p = 0.98) and in the heart and brain frozen for 3 months (p = 0.07 and 0.28, respectively) or more (p = 0.21 for heart and p > 0.99 for brain). In addition, CAT and SOD activities were reduced by storage duration in all tissues evaluated (p < 0.05). Our findings show that sample storage duration alters the quantification of oxidative stress markers in mice submitted to maximum exercise, and its effect is tissue and marker dependent. Some recommendations to achieve more accurate and reproducible data in the exercise physiology and oxidative stress markers field are presented.

[Quality of life of patients with colorectal cancer receiving outpatient chemotherapy]. / Qualidade de vida do paciente com câncer colorretal em quimioterapia ambulatorial.

This is a cross-sectional study with a descriptive quantitative approach, which aimed to assess the quality of life (QOL) of patients with colorectal cancer receiving outpatient chemotherapy. The research was conducted in an Outpatient Chemotherapy Unit at a hospital in southern Brazil, whose patients were diagnosed with colorectal cancer and were treated with the 5-FU protocol. The sample had 48 participants who were undergoing chemotherapy for a period of six months. A questionnaire, the WHOQOL-Bref was used as an instrument. In the results, the age of 50 years or more with at least a month and a maximum of 11 months of treatment prevailed. The domains of the WHOQOL-Bref more significantly affected were the psychological and the social relations one, respectively, with significant differences in responses regarding overall QOL in those who were in the first cycle of treatment from those already in the 6th cycle.

An Intervention to Increase Situational Awareness and the Culture of Mutual Care (Foco) and Its Effects During COVID-19 Pandemic: A Randomized Controlled Trial and Qualitative Analysis.

Situational awareness is especially important to decision-making in health care. Comprehending the situation is crucial for anticipating any change in the environment and delivering optimal care. The objective of this study was to evaluate the effects of a training to increase situational awareness and mutual care designed for health care workers (FoCo) in a randomized controlled trial with additional qualitative analysis. We also investigated the perception of the training for the COVID-19 pandemic moment, in May 2020, almost 6 months after we finished the data collection at the Emergency Care Unit, which became a COVID-19 treatment reference for the care of a population depending on the public health system, in Sao Paulo, Brazil. We conclude that FoCo training can be an important instrument for health care professionals both in times of pandemic and "normal times," to increase situational awareness, the culture of mutual care and decrease the possibility of occupational injuries and illnesses.

Accuracy of the clinical indicators of ineffective health management in celiac people.

OBJECTIVE: to analyze the accuracy of clinical indicators of "Ineffective health management" in celiac patients and to verify associations between sociodemographic characteristics and clinical indicators. METHOD: a cross-sectional study, conducted from May to September 2017, with 83 celiac patients, through an interview. Accuracy measures were defined by latent class model. RESULTS: there was a prevalence of "Ineffective health management" of 55.69%. "Failure to take action to reduce risk factor" and "Failure to include treatment regimen in daily living" better predict this diagnosis. Paid occupation reduces the chance of the presence of "Difficulty with prescribed regimen". Participation in support association reduces the chance of the presence of "Difficulty with prescribed regimen", "Ineffective choices in daily living for meeting health goal" and "Failure to take action to reduce risk factor". CONCLUSION: accurate clinical indicators identification assists clinical reasoning for diagnostic inference in specific health contexts.

Human pluripotent stem cell-derived cardiomyocytes as a target platform for paracrine protection by cardiac mesenchymal stromal cells.

Ischemic heart disease remains the foremost cause of death globally, with survivors at risk for subsequent heart failure. Paradoxically, cell therapies to offset cardiomyocyte loss after ischemic injury improve long-term cardiac function despite a lack of durable engraftment. An evolving consensus, inferred preponderantly from non-human models, is that transplanted cells benefit the heart via early paracrine signals. Here, we tested the impact of paracrine signals on human cardiomyocytes, using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) as the target of mouse and human cardiac mesenchymal stromal cells (cMSC) with progenitor-like features. In co-culture and conditioned medium studies, cMSCs markedly inhibited human cardiomyocyte death. Little or no protection was conferred by mouse tail tip or human skin fibroblasts. Consistent with the results of transcriptomic profiling, functional analyses showed that the cMSC secretome suppressed apoptosis and preserved cardiac mitochondrial transmembrane potential. Protection was independent of exosomes under the conditions tested. In mice, injecting cMSC-conditioned media into the infarct border zone reduced apoptotic cardiomyocytes > 70% locally. Thus, hPSC-CMs provide an auspicious, relevant human platform to investigate extracellular signals for cardiac muscle survival, substantiating human cardioprotection by cMSCs, and suggesting the cMSC secretome or its components as potential cell-free therapeutic products.

Loss of MafA and MafB expression promotes islet inflammation.

Maf transcription factors are critical regulators of beta-cell function. We have previously shown that reduced MafA expression in human and mouse islets is associated with a pro-inflammatory gene signature. Here, we investigate if the loss of Maf transcription factors induced autoimmune processes in the pancreas. Transcriptomics analysis showed expression of pro-inflammatory as well as immune cell marker genes. However, clusters of CD4+ T and B220+ B cells were associated primarily with adult MafA-/-MafB+/-, but not MafA-/- islets. MafA expression was detected in the thymus, lymph nodes and bone marrow suggesting a novel role of MafA in regulating immune-cell function. Analysis of pancreatic lymph node cells showed activation of CD4+ T cells, but lack of CD8+ T cell activation which also coincided with an enrichment of naïve CD8+ T cells. Further analysis of T cell marker genes revealed a reduction of T cell receptor signaling gene expression in CD8, but not in CD4+ T cells, which was accompanied with a defect in early T cell receptor signaling in mutant CD8+ T cells. These results suggest that loss of MafA impairs both beta- and T cell function affecting the balance of peripheral immune responses against islet autoantigens, resulting in local inflammation in pancreatic islets.

Single Cell Gene Expression to Understand the Dynamic Architecture of the Heart.

The recent development of single cell gene expression technologies, and especially single cell transcriptomics, have revolutionized the way biologists and clinicians investigate organs and organisms, allowing an unprecedented level of resolution to the description of cell demographics in both healthy and diseased states. Single cell transcriptomics provide information on prevalence, heterogeneity, and gene co-expression at the individual cell level. This enables a cell-centric outlook to define intracellular gene regulatory networks and to bridge toward the definition of intercellular pathways otherwise masked in bulk analysis. The technologies have developed at a fast pace producing a multitude of different approaches, with several alternatives to choose from at any step, including single cell isolation and capturing, lysis, RNA reverse transcription and cDNA amplification, library preparation, sequencing, and computational analyses. Here, we provide guidelines for the experimental design of single cell RNA sequencing experiments, exploring the current options for the crucial steps. Furthermore, we provide a complete overview of the typical data analysis workflow, from handling the raw sequencing data to making biological inferences. Significantly, advancements in single cell transcriptomics have already contributed to outstanding exploratory and functional studies of cardiac development and disease models, as summarized in this review. In conclusion, we discuss achievable outcomes of single cell transcriptomics' applications in addressing unanswered questions and influencing future cardiac clinical applications.

Anisakidae parasites in frozen fish fillets intended for human consumption / Parásitos anisákidos en filetes de pescado congelado destinados a consumo humano

Introduction: Anisakiasis is a human parasitic disease caused by the consumption of raw fish or shellfish containing larvae of the Anisakidae family. It is currently considered an emerging disease of public health interest. Objective: To identify the presence of larvae of the Anisakidae family in samples of frozen raw fish fillets intended for human consumption in markets in Medellín and its metropolitan area in Antioquia, Colombia. Materials and methods: A cross-sectional study was carried out, in which larvae of the Anisakidae family were detected and identified in frozen raw fish fillets from three representative markets in Medellín and its metropolitan area. A total of 384 ready for consumption fillets were analyzed (197 sawfish, 137 salmon, 37 tuna, and 13 hake), using the pressing and ultraviolet light method. Taxonomic keys were used to identify the collected parasites and to establish its genus. Conventional PCR and Sanger sequencing was performed to determine the species. Results: Four larvae were found in 4 of the 384 (1.04%) fillets (CI95% 1.04 ± 1.01%). The species of fish in which the larvae were found was sawfish (Scomberomorus spp.) and the genus and species of the larvae was established as Anisakis pegreffii. Conclusions: According to the study, the presence of Anisakis parasites in frozen raw fish fillets in the influence area is evident.

Introducción. La anisakiasis es una infección producida por parásitos de la familia Anisakidae, transmitida a los humanos por el consumo de pescado o mariscos crudos. En la actualidad, se considera una enfermedad emergente de interés en salud pública. Objetivo. Identificar la presencia de larvas de la familia Anisakidae en muestras de filetes de pescado crudo congelado destinados a consumo humano, en mercados de Medellín y su área metropolitana en Antioquia (Colombia). Materiales y métodos. Se realizó un estudio transversal, en el cual se buscó la presencia de larvas de la familia Anisakidae en filetes de pescado crudo congelado de tres mercados representativos de Medellín y su área metropolitana. Se analizaron 384 filetes listos para el consumo (197 sierras, 137 salmones, 37 atunes y 13 merluzas). Cada filete fue analizado mediante el método de prensado y luz ultravioleta. Los parásitos recolectados se identificaron a partir de claves taxonómicas para establecer el género, así como PCR convencional y posterior secuenciación Sanger, para determinar la especie. Resultados. Se encontraron 4 larvas en 4 de los 384 filetes (1,04 %) (IC95% 1,04 ± 1,01 %). Las larvas encontradas fueron identificadas como Anisakis pegreffi y el tipo de pescado en el cual se encontraron fue la sierra (Scomberomorus spp.) Conclusiones. De acuerdo con el estudio realizado, se evidencia la presencia de parásitos anisákidos en filetes de pescado crudo congelado en el área de influencia.

Ontogenetic development of the inner ear saccule and utricle in the Lusitanian toadfish: Potential implications for auditory sensitivity.

Studies addressing structure-function relationships of the fish auditory system during development are sparse compared to other taxa. The Batrachoididae has become an important group to investigate mechanisms of auditory plasticity and evolution of auditory-vocal systems. A recent study reported ontogenetic improvements in the inner ear saccule sensitivity of the Lusitanian toadfish, Halobatrachus didactylus, but whether this results from changes in the sensory morphology remains unknown. We investigated how the macula and organization of auditory receptors in the saccule and utricle change during growth in this species. Inner ear sensory epithelia were removed from the end organs of previously PFA-fixed specimens, from non-vocal posthatch fry (<1.4 cm, standard length) to adults (>23 cm). Epithelia were phalloidin-stained and analysed for area, shape, number and orientation patterns of hair cells (HC), and number and size of saccular supporting cells (SC). Saccular macula area expanded 41x in total, and significantly more (relative to body length) among vocal juveniles (2.3-2.9 cm). Saccular HC number increased 25x but HC density decreased, suggesting that HC addition is slower relative to epithelial growth. While SC density decreased, SC apical area increased, contributing to the epithelial expansion. The utricule revealed increased HC density (striolar region) and less epithelial expansion (5x) with growth, contrasting with the saccule that may have a different developmental pattern due to its larger size and main auditory functions. Both macula shape and HC orientation patterns were already established in the posthatch fry and retained throughout growth in both end organs. We suggest that previously reported ontogenetic improvements in saccular sensitivity might be associated with changes in HC number (not density), size and/or molecular mechanisms controlling HC sensitivity. This is one of the first studies investigating the ontogenetic development of the saccule and utricle in a vocal fish and how it potentially relates to auditory enhancement for acoustic communication.

Term amniotic fluid: an unexploited reserve of mesenchymal stromal cells for reprogramming and potential cell therapy applications.

BACKGROUND: Mesenchymal stromal cells (MSCs) are currently being evaluated in numerous pre-clinical and clinical cell-based therapy studies. Furthermore, there is an increasing interest in exploring alternative uses of these cells in disease modelling, pharmaceutical screening, and regenerative medicine by applying reprogramming technologies. However, the limited availability of MSCs from various sources restricts their use. Term amniotic fluid has been proposed as an alternative source of MSCs. Previously, only low volumes of term fluid and its cellular constituents have been collected, and current knowledge of the MSCs derived from this fluid is limited. In this study, we collected amniotic fluid at term using a novel collection system and evaluated amniotic fluid MSC content and their characteristics, including their feasibility to undergo cellular reprogramming. METHODS: Amniotic fluid was collected at term caesarean section deliveries using a closed catheter-based system. Following fluid processing, amniotic fluid was assessed for cellularity, MSC frequency, in-vitro proliferation, surface phenotype, differentiation, and gene expression characteristics. Cells were also reprogrammed to the pluripotent stem cell state and differentiated towards neural and haematopoietic lineages. RESULTS: The average volume of term amniotic fluid collected was approximately 0.4 litres per donor, containing an average of 7 million viable mononuclear cells per litre, and a CFU-F content of 15 per 100,000 MNCs. Expanded CFU-F cultures showed similar surface phenotype, differentiation potential, and gene expression characteristics to MSCs isolated from traditional sources, and showed extensive expansion potential and rapid doubling times. Given the high proliferation rates of these neonatal source cells, we assessed them in a reprogramming application, where the derived induced pluripotent stem cells showed multigerm layer lineage differentiation potential. CONCLUSIONS: The potentially large donor base from caesarean section deliveries, the high yield of term amniotic fluid MSCs obtainable, the properties of the MSCs identified, and the suitability of the cells to be reprogrammed into the pluripotent state demonstrated these cells to be a promising and plentiful resource for further evaluation in bio-banking, cell therapy, disease modelling, and regenerative medicine applications.