Human Tissue Research Research Articles

New self-assembling multifunctional templates for the biofabrication and controlled self-release of cultured tissue.

The need to source live human tissues for research and clinical applications has been a major driving force for the development of new biomaterials. Ideally, these should elicit the formation of scaffold-free tissues with native-like structure and composition. In this study, we describe a biologically interactive coating that combines the fabrication and subsequent self-release of live purposeful tissues using template–cell–environment feedback. This smart coating was formed from a self-assembling peptide amphiphile comprising a protease-cleavable sequence contiguous with a cell attachment and signaling motif. This multifunctional material was subsequently used not only to instruct human corneal or skin fibroblasts to adhere and deposit discreet multiple layers of native extracellular matrix but also to govern their own self-directed release from the template solely through the action of endogenous metalloproteases. Tissues recovered through this physiologically relevant process were carrier-free and structurally and phenotypically equivalent to their natural counterparts. This technology contributes to a new paradigm in regenerative medicine, whereby materials are able to actively direct and respond to cell behavior. The novel application of such materials as a coating capable of directing the formation and detachment of complex tissues solely under physiological conditions can have broad use for fundamental research and in future cell and tissue therapies.

The reporting of research ethics committee approval and informed consent in otolaryngology journals.

Medical research involving human subjects must follow ethical standards as outlined in the Declaration of Helsinki of the World Medical Association. The aim of this study was to assess the frequency of reporting of informed consent and regional ethical committee (REC) approval in all reports of trials published in the major European Otolaryngology journals. Review of all clinical research articles published online in the calendar year 2012. Three leading European Otolaryngology journals. Clinical Otolaryngology, The Journal of Laryngology and Otology and The European Achieves of Oto-Rhino-Laryngology. Evaluate the incidence of reporting of REC approval and informed consent. Of the 767 articles reviewed, 401 met the inclusion criteria (manuscripts reporting human subjects, human tissue or identifiable personal data research which require ethical approval). 49.9% lacked a statement of REC approval and 42.9% lacked disclosure of informed consent. Articles that did not state REC approval were associated with not stating informed consent (P<0.05). Articles that lack explicit statements of REC approval and informed consent are frequent and continue to be published in major otolaryngology journals.

Cadaver Dissection in Anatomy: The Ethical Aspect

Cadaveric dissection is an integral part of medical education and the basis of anatomy for first M.B.B.S. students. The dead human body has failed to elicit much theological interest, and until recently, little ethical interest. Dissection of cadavers is a time honored part of medical education. Undergraduate and postgraduate students do dissection as a part of learning human anatomy. Cadavers are obtained from the forensic mortuary declared as ‘unclaimed body’ or through donated bodies that includes voluntary donation during life or their after by the relatives. However, use of human tissue for research and the use of human cadavers for teaching and training purposes are surrounded by ethical issues. In this paper we will be discussing the ethical issues in relation to the use of cadaver i.e. respect for human body even after life.

SIVQ-LCM Protocol for the ArcturusXT Instrument

SIVQ-LCM is a new methodology that automates and streamlines the more traditional, user-dependent laser dissection process. It aims to create an advanced, rapidly customizable laser dissection platform technology. In this report, we describe the integration of the image analysis software Spatially Invariant Vector Quantization (SIVQ) onto the ArcturusXT instrument. The ArcturusXT system contains both an infrared (IR) and ultraviolet (UV) laser, allowing for specific cell or large area dissections. The principal goal is to improve the speed, accuracy, and reproducibility of the laser dissection to increase sample throughput. This novel approach facilitates microdissection of both animal and human tissues in research and clinical workflows.

SIVQ-LCM protocol for the ArcturusXT instrument.

SIVQ-LCM is a new methodology that automates and streamlines the more traditional, user-dependent laser dissection process.It aims to create an advanced, rapidly customizable laser dissection platform technology. In this report, we describe the integration of the image analysis software Spatially Invariant Vector Quantization (SIVQ) onto the ArcturusXT instrument. The ArcturusXT system contains both an infrared (IR) and ultraviolet (UV) laser, allowing for specific cell or large area dissections. The principal goal is to improve the speed, accuracy, and reproducibility of the laser dissection to increase sample throughput. This novel approach facilitates microdissection of both animal and human tissues in research and clinical workflows.

Tissue banking in ENT: challenges and methods

Biobanking is the process of storing high quality human biospecimens alongside linked clinical data, for research purposes. The aim is to identify novel biomarkers with prognostic or diagnostic significance. However, the challenges implicit in the collection and storage of human tissue for research have curtailed the impact of this technique to date. This paper aims to summarise the challenges faced by biobanking within the ENT specialty in the UK, and to present protocols used for the routine collection, freezing and storage of tissue specimens at the Royal National Throat, Nose and Ear Hospital. These protocols could be used to guide other ENT departments (in the UK and worldwide) wishing to initiate the routine collection and storage of tissue samples. Their publication could also help to establish basic standards and ensure consistency in ENT tissue storage. Interviews conducted with industry experts, and a literature review of 'best practice' in biobanking. The ENT specialty must stay abreast of progress in human tissue research in order to ensure the best possible management of its patients. Our protocol for the routine banking of ENT tissue at the Royal National Throat, Nose and Ear Hospital could be used as a template for other ENT departments (in the UK and worldwide) to encourage widespread implementation of high quality tissue banking.

Impossible, impractical, and non-identifiable? New criteria regarding consent for human tissue research in the Declaration of Helsinki.

The 2008 revision of the Declaration of Helsinki (Declaration) includes a new paragraph dealing specifically with the standards of informed consent required for research involving identifiable human tissue samples and/or data. In cases where obtaining consent would be impossible or unduly burdensome, researchers may now proceed without it, following approval of the project by an ethics committee. This is a significant development in the Declaration, yet so far it has received little attention. We examine the implications of paragraph 25, and assess its role in the debates on proper sample handling. In particular, we question whether the use of the term "identifiable" weakens the paragraph, as its meaning depends on national context. Relying on this term to designate samples that could be traced to the donor, and therefore carry risk for that donor, is impossible if its meaning is not universally accepted. The Declaration of Helsinki is now entering a new revision phase. In order to protect sample donors, paragraph 25 should be enhanced, and the remit of the Declaration strengthened, by a more precise description of which samples and data count as identifiable.

Rewards and incentives for the provision of human tissue for research

The Nuffield Council on Bioethics’ 2011 report, Human Bodies: Donation for Medicine and Research, proposes a system for examining the ethical implications of different types of incentives for the provision...

Engineering Freedom? A Critique of Biotechnological Routes to Animal Liberation

This essay presents a critical analysis of the claims of three biotechnological innovations to be facilitative of nonhuman animal liberation: de-domestication; in vitro meat (IVM); and human tissue research. The analysis is facilitated by the application of a conceptual model of human/nonhuman animal relations, which highlights the interconnection of hegemonic material and discursive practices in classifying the nature and purpose of nonhuman animals. The analysis therefore considers both the liberatory potential of each biotechnology, as well as its limitations, in light of its capacity to, respectively, generate novel material and discursive practices or to reproduce existing ones. The essay concludes with an extended discussion of an alternative, nonbiotechnological approach to animal liberation rooted in the ecofeminist work of Marti Kheel and Brian Luke.

Biobanking for research in surgery: are surgeons in charge for advancing translational research or mere assistants in biomaterial and data preservation?

High-quality biospecimens of human origin with annotated clinical and procedural data are an important tool for biomedical research, not only to map physiology, pathophysiology and aetiology but also to go beyond in translational research. This has opened a new special field of research known as 'biobanking', which focuses on how to collect, store and provide these specimens and data, and which is substantially supported by national and European funding. An overview on biobanking is given, with a closer look on a clinical setting, concerning a necessary distinction from clinical trials and studies as well as a comparison of prospective sample collection with secondary use of archived samples from diagnostics. Based on a summary of possible use and scientific impact of human tissue in research, it is shown how surgical expertise boosts the scientific value of specimens and data. Finally, an assessment of legal and ethical issues especially from a surgical perspective is given, followed by a model of interdisciplinary biobanking within a joint 'centre' that as synergistic structure merges essential input from surgery as well as laboratory medicine, pathology and biometry. Within the domain of biobanking, surgeons have to develop a better awareness of their role within translational research, not only on the level of medical faculties but also as nationally and internationally funded initiatives. Therefore, the authors suggest a platform for biobanking within the German association of surgeons in analogy to the existing special interest group for clinical trials.

Translational neuropharmacology: the use of human isolated gastrointestinal tissues

Translational sciences increasingly emphasize the measurement of functions in native human tissues. However, such studies must confront variations in patient age, gender, genetic background and disease. Here, these are discussed with reference to neuromuscular and neurosecretory functions of the human gastrointestinal (GI) tract. Tissues are obtained after informed consent, in collaboration with surgeons (surgical techniques help minimize variables) and pathologists. Given the difficulties of directly recording from human myenteric neurones (embedded between muscle layers), enteric motor nerve functions are studied by measuring muscle contractions/relaxations evoked by electrical stimulation of intrinsic nerves; responses are regionally dependent, often involving cholinergic and nitrergic phenotypes. Enteric sensory functions can be studied by evoking the peristaltic reflex, involving enteric sensory and motor nerves, but this has rarely been achieved. As submucosal neurones are more accessible (after removing the mucosa), direct neuronal recordings are possible. Neurosecretory functions are studied by measuring changes in short-circuit current across the mucosa. For all experiments, basic questions must be addressed. Because tissues are from patients, what are the controls and the influence of disease? How long does it take before function fully recovers? What is the impact of age- and gender-related differences? What is the optimal sample size? Addressing these and other questions minimizes variability and raises the scientific credibility of human tissue research. Such studies also reduce animal use. Further, the many differences between animal and human GI functions also means that human tissue research must question the ethical validity of using strains of animals with unproved translational significance.

Re-moralising medicine: The bioethical thought collective and the regulation of the body in British medical research

The way in which the scientific and medical use of the human body is problematised and governed in the United Kingdom was radically reconfigured over the last 30 years, changing from a logic of rule articulated around ‘supply’ and ‘solidarity’ to one construed around ‘ethics’. Drawing on the work of Ludwik Fleck and others, this article argues that one of the reasons for this reconfiguration was the existence and influence of a network of philosophers, doctors and lawyers who sought, from the 1960s onwards, to re-moralise medicine: the bioethical thought collective. The article first describes the collective’s membership and organisation, focusing in particular on the form of the interdisciplinary expert committee. It also describes some of the knowledge and practices that make up the community’s thought style, such as its moral concern about modern medicine and the notions of respect for persons and informed consent. The article then shows how these organisational forms, knowledge and practices that characterise the collective have shaped the government of human tissue research over the last 15 years. By highlighting the important role played by expert networks and knowledge, the article makes an original contribution to the sociology of the ethical government of biomedical science.

Generation and characterization of new HER2 monoclonal antibodies

Antibodies are among the most commonly used research and diagnostic tools. Antibody type and clonality are important in any assay as they can influence epitope detection. HER2 oncoprotein is overexpressed or undergoes gene amplification in approximately 30% of invasive breast carcinomas and 20% of gastric adenocarcinomas. Overexpression of HER2 is primarily detected by immunohistochemistry (IHC) on neoplastic tissue sections. We produced five murine hybridoma clones secreting monoclonal antibodies (MAbs) against HER2 protein. For hybridoma production, spleen cells from BALB/c mice immunized with a recombinant fragment of the extracellular portion of HER2 (rHER2) were fused to SP2/O-Ag14 cells, selected in HAT medium and screened by indirect ELISA. MAbs secreted were characterized according to isotypes, functional affinity constants, reaction with the native protein in MCF-7 cells by indirect immunofluorescence and in tissue sections from HER2 positive breast cancer specimens by IHC. Two MAbs were IgG2b and three were IgG1, and their affinity constants ranged from 6×107 to 1×109M−1. All MAbs reacted with the native protein and two stained strongly the membrane of neoplastic cells overexpressing HER2. These two MAbs could be useful in assaying HER2 overexpression in human tissues for research and possibly diagnostic purposes after a proper large-scale validation study.

Human Tissue Research: A European Perspective on Ethical &amp; Legal Challenges

Human Tissue Research: A European Perspective on Ethical &amp; Legal Challenges

P2‐404: Alzheimer's disease and the Valley of Death: Not enough guidance from human brain tissue?

Medical science is currently perceived as underperforming. This is because of the relatively slow recent rate of development of new disease treatments. This has been blamed on cultural, regulatory, and economic factors that generate a so-called Valley of Death, hindering new drug candidates from being moved into clinical trials and eventually approved for use. We propose, however, that for neurodegenerative diseases, a relative decline of human brain tissue research is also a contributor. The present pharmacological agents for treating Alzheimer's disease (AD) were identified through direct examination of postmortem human brain tissue more than 30 years ago. Since that time the percentage of research grants awarded to human brain tissue-using projects has dropped precipitously and publication rates have stagnated. As human brain tissue research has played a central and often initiating role in identifying most of the targets that have gone to AD clinical trials, it is proposed that the rate of discovery of new targets has been curtailed. Additionally, the continued rejection of cortical biopsy as a diagnostic method for AD has most probably depressed the perceived effect sizes of new medications and contributed to the high Phase II clinical trial failure rates. Despite the relative lack of funding, human brain discovery research has continued to make important contributions to our understanding of neurodegenerative disease, and brain banks have played an essential role. It is likely that the pace of discovery will dramatically accelerate over the coming decades as increasingly powerful tools including genomics, epigenetics, transcriptomics, regulatory RNA, gene expression profiling, proteomics, and metabolomics are applied. To optimize the promise of these new technologies, however, it is critical that brain banks are rejuvenated by enhanced governmental and/or private support.

Ethics in dissection of cadaver in teaching and learning of anatomy

Dissection of dead body is a time honoured part of medical education. Undergraduate and postgraduate students do dissection as a part of the learning human anatomy. According to the state regulations, most of the cadavers are obtained from the forensic mortuary declared as ‘unclaimed body’. Some are collected through ‘body donation’ as well. However, use of human tissue for research and the use of human cadavers for teaching and training purposes are surrounded by ethical uncertainties. Moreover, the main ethical concern of cadaver dissection lies in respect to human life. This writing has been aimed to put a nominal guideline how to act in an ethical way in cadaver dissection by the medical students in medical colleges and pave the way for better understanding how to respond in an ethical manner in medical professional life. Some rules and regulations have been recommended for maintaining the highest standard of a dissection hall as well as the dignity of the cadavers. DOI: http://dx.doi.org/10.3329/bioethics.v2i3.10258 Bangladesh Journal of Bioethics 2011;2(3):10-15

Toward Clarity in the Use of Human Tissue in Research

Toward Clarity in the Use of Human Tissue in Research

A Critical Need for Increased Understanding of the Use of Human Tissues in Biomedical Research

The recent statement, “only a small fraction of the samples being submitted (for the Cancer Genome Atlas [TCGA] project) were of suitable quality for research,” in an article in Genome Technology [1] caught my attention because it is an example of a general lack of understanding of specific issues regarding the provision of human tissues to support biomedical research [2]. The implication of this statement is that because tissues do not meet very demanding requirements, the tissues were of poor quality. I will use this statement to present examples of problems in understanding the use of human tissues in research. In this case, “quality” was confused with “availability” – one among several common misconceptions among investigators.

Adiponectin upregulates hepatocyte CMKLR1 which is reduced in human fatty liver

Objective: Chemokine-like receptor 1 (CMKLR1) ligands chemerin and resolvin E1 are suggested to have a role in non-alcoholic fatty liver disease (NAFLD). Here, expression of CMKLR1 in liver cells and NAFLD was studied. Methods: Primary human hepatocytes and liver of patients with and without liver steatosis were used. Human liver tissue was obtained according to the guidelines of the charitable state-controlled foundation Human Tissue & Cell Research (HTCR) with the patient's informed consent. To induce NASH mice were fed a methionine-choline deficient diet for 6 weeks. Results: CMKLR1 was detected in primary human hepatocytes (PHH), Kupffer cells, bile-duct cells and hepatic stellate cells. In human fatty liver and in fibrotic liver of mice fed a methionine-choline deficient diet CMKLR1 protein was reduced. Hepatocytes are the major cells in the liver and effects of adipokines, cytokines and lipids on CMKLR1 in PHH were analysed. Increased cellular triglyceride or cholesterol content, lipopolysaccharide, IL–6, TNF and leptin did not influence CMKLR1 levels in PHH whereas profibrotic TGFbeta tended to reduce CMKLR1. Adiponectin strongly upregulated CMKLR1 mRNA and protein, and CMKLR1 was suppressed in the liver of adiponectin deficient mice. Conclusion: These data indicate that low CMKLR1 in NAFLD may partly result from reduced adiponectin activity.

Translational Research in Pediatrics: Tissue Sampling and Biobanking

Translational research is expanding and has become a focus of National Research funding agencies, touted as the primary avenue to improve health care practice. The use of human tissues for research on disease etiology is a pillar of translational research, particularly with innovations in research technologies to investigate the building blocks of disease. In pediatrics, translational research using human tissues has been hindered by the many practical and ethical considerations associated with tissue procurement from children and also by a limited population base for study, by the increasing complexities in conducting clinical research, and by a lack of dedicated child-health research funding. Given these obstacles, pediatric translational research can be enhanced by developing strategic and efficient biobanks that will provide scientists with quality tissue specimens to render accurate and reproducible research results. Indeed, tissue sampling and biobanking within pediatric academic settings has potential to impact child health by promoting bidirectional interaction between clinicians and scientists, helping to maximize research productivity, and providing a competitive edge for attracting and maintaining high-quality personnel. The authors of this review outline key issues and practical solutions to optimize pediatric tissue sampling and biobanking for translational research, activities that will ultimately reduce the burden of childhood disease.