Overview of a quality assurance/quality control compliance program consistent with FDA regulations and policies for somatic cell and gene therapies: a four year experience

Somatic cell and gene therapy involve the application of biological technologies to an individual patient through the use of living cells which provide a therapeutic benefit (Aliski, 1991). Various forms of cellular and gene therapies are being developed and evaluated in an increasing number of clinical trials for congential and acquired disorders. The potential and progress of these therapeutic applications have resulted in an increasing effort by the Food and Drug Administration (FDA) to develop the regulatory framework under which these therapeutic approaches would insure safety and efficacy, the primary mandate of the FDA.Over five years ago Cellcor began to define the parameters, specifications, and conditions relevant to a Quality Assurance/Quality Control (QA/QC) program that has evolved to insure safety and maximize the efficacy of applications of the company'sex vivo technology, autolymphocyte therapy. Autolymphocyte therapy is an outpatient form of somatic cell immunotherapy based upon the infusion of T cells that have been activatedex vivo using a combination of previously generated autologous cytokines and an anti-CD3 monoclonal antibody.We have been able to demonstrate the feasibility for the safe, controlled, and consistent preparation and delivery of a cellular therapy by application of relevant GMP regulations. This presentation reviews aspects of this program and chronicles our experience which at present amounts to over 4400 infusions for over 700 patients. This program provides a high degree of assurance that a cellular therapy program can be carried out in a multisite mode involving hundreds of patients through the strict adherence to cGMP as set forth in existing regulations. It would be prudent that developers of cellular andex vivo gene therapies establish a similar cell processing and QA/QC infrastructure at an early developmental stage to optimize safety and reproducibility and facilitate regulatory review.     

POSTOPERATIVE-POSTANESTHETIC RECOVERY ROOMS—A Report on Three Years of Experience

An analysis of the three-year experience of the recovery room at the Cedars of Lebanon Hospital emphasized the importance of this phase of postoperative care. Closer observation and supervision of the patient is provided during a critical period, and specialized nursing care is constantly available. Supportive and corrective therapy can be carried out immediately by the cooperative efforts of the anesthesiologist and surgeon, who are close by. The completely equipped and properly staffed recovery room is the best assurance for reducing postoperative morbidity and mortality.     

Postoperative-postanesthetic recovery rooms; a report on three years of experience

An analysis of the three-year experience of the recovery room at the Cedars of Lebanon Hospital emphasized the importance of this phase of postoperative care. Closer observation and supervision of the patient is provided during a critical period, and specialized nursing care is constantly available. Supportive and corrective therapy can be carried out immediately by the cooperative efforts of the anesthesiologist and surgeon, who are close by. The completely equipped and properly staffed recovery room is the best assurance for reducing postoperative morbidity and mortality.     

细胞药物的标准及质量控制——细胞药物连载之四

细胞药物最终用于人体,必须建立相应的质量标准,进行质量控制。系统地贯彻到供者筛查、组织采集、细胞分离、培养、冻存、复苏、放行、运输、使用等全过程,确保产品的安全性、有效性和稳定性。近年来,我国逐渐改变了把细胞治疗作为第三类医疗技术管理的思路。一方面,已有第三类医疗技术取消行政审批;另一方面,又把除自体外的干细胞移植纳入药物管理,并建立了相应的质量标准和质量管理办法。     

Evaluation of Early Diagnostic Services for the Elderly

Medical examination was offered to a group of “high risk” old people who were not necessarily patients or known to their family doctors, but with the agreement of these family doctors. Two clinics set up for this purpose have been running for several years, and the results of examination and follow-up of 300 consecutive patients are reported.Major conditions were found in two-thirds of patients producing functional impairment in most of these.Recommendations as to therapy and management were carried out in 161 of 194 patients but not in the remainder.Clear evidence of improvement was found in half of the patients who carried out recommendations, and this improvement was attributable to earlier diagnosis than would have been achieved without these clinics in 42% of cases.Including all patients examined, the proportion helped by early diagnosis at 18 to 30 months'' follow-up was 23%.It is concluded that the offer of a routine examination to high risk groups is of benefit to old people and a form of medical practice which should be widely adopted.     

Host tissue response in stem cell therapy

Preclinical and clinical trials of stem cell therapy have been carried out for treating a broad spectrum of diseases using several types of adult stem cells. While encouraging therapeutic results have been obtained, much remains to be investigated regarding the best cell type to use, cell dosage, delivery route, long-term safety, clinical feasibility, and ultimately treatment cost. Logistic aspects of stem cell therapeutics remain an area that requires urgent attention from the medical community. Recent cardiovascular trial studies have demonstrated that growth factors and cytokines derived from the injected stem cells and host tissue appear to contribute largely to the observed therapeutic benefits, indicating that trophic actions rather than the multilineage potential (or stemness) of the administered stem cells may provide the underlying tissue healing power. However, the capacity for trophic factor production can be aberrantly downregulated as seen in human heart disease. Skeletal muscle is a dynamic tissue with an impressive ability to continuously respond to environmental stimuli. Indeed, a relation exists between active skeletal muscle and low cardiovascular risk, highlighting the critical link between the skeletal muscle and cardiovascular systems. Adding to this notion are recent studies showing that stem cells injected into skeletal muscle can rescue the failing rodent heart through activation of the muscle trophic factor network and mobilization of bone marrow multilineage progenitor cells. However, aging and disease can adversely affect the host tissue into which stem cells are injected. A better understanding of the host tissue response in stem cell therapy is necessary to advance the field and bridge the gap between preclinical and clinical findings.     

A 3-year experience of quality control and quality assurance in the multisite delivery of a lymphocyte-based cellular therapy for renal cell carcinoma

Autolymphocyte therapy (ALT) is outpatient-based adoptive immunotherapy using ex vivo-activated memory T-cells. To support the safe and reproducible delivery of ALT at three cell processing facilities (Boston, MA; Atlanta, GA; Orange, CA) we created a comprehensive quality assurance/quality control program compliant with recent FDA guidance relevant to activated lymphocytes and somatic cell therapies. Each facility performed extensive QC testing to ensure sterility, viability, and proper cell yield. Additonally, several QC tests were performed at Cellocr's centralized reference laboratory to monitor cell potency and identity of the ex vivo-processed lymphocytes. We report here the successful implementation of this QA/QC program for ALT which has resulted in the safe preparation and delivery of cell infusion products amounting to over 3600 treatments at seven clinical sites nationwide. We believe this program will serve as a model for other cellular therapies.     

基于LED作为照射光源的血疗法可行性研究

通过对LED在生物医学领域的应用分析发现,LED光存在与激光相类似的生物刺激效应。目前弱激光血疗已由血管内照射发展到体表照射、黏膜照射。这为LED光照射体表、黏膜实现血疗提供了良好的预期。为了证明LED光可以代替激光作为净血治疗仪的光源,本实验设计了激光与LED光对自由基损伤模型红细胞变形性作用的对比实验,结果表明,两种光照方法均对红细胞变形性有改善作用。LED相比激光安全性更好、成本更加低廉,特别适合往家用小型化方向发展。     

Governing stem cell therapy in India: regulatory vacuum or jurisdictional ambiguity?

Stem cell treatments are being offered in Indian clinics although preclinical evidence of their efficacy and safety is lacking. This is attributed to a governance vacuum created by the lack of legally binding research guidelines. By contrast, this paper highlights jurisdictional ambiguities arising from trying to regulate stem cell therapy under the auspices of research guidelines when treatments are offered in a private market disconnected from clinical trials. While statutory laws have been strengthened in 2014, prospects for their implementation remain weak, given embedded challenges of putting healthcare laws and professional codes into practice. Finally, attending to the capacities of consumer law and civil society activism to remedy the problem of unregulated treatments, the paper finds that the very definition of a governance vacuum needs to be reframed to clarify whose rights to health care are threatened by the proliferation of commercial treatments and individualized negligence-based remedies for grievances.     

关节腔内注射细胞制剂治疗骨关节炎的临床研究进展

关节腔内注射细胞制剂是治疗骨关节炎(OA)的新趋势,且展现了良好的临床效果、安全性及可行性。细胞制剂有多种类型,包括血小板、干细胞和单个核细胞(MNCs)。血小板制剂倾向于基层医疗机构应用和早期OA患者的治疗;MNCs较适用于不具备干细胞培养条件的医疗机构和中期OA患者应用;而干细胞制剂较适合高级别医疗机构和晚期OA患者应用。虽然腔内注射细胞产品治疗OA进行了大量临床研究,但亦有不足;因此制定腔内注射治疗OA不同细胞制剂的质量控制标准、临床应用标准和进行大样本多中心随机对照临床试验势在必行。探索外泌体、外周血MNCs治疗OA的临床研究及其分子机制,是研究的新切入点。     

基于区域协同医疗的检测技术需求分析与解决方案

在分析县级医院及基层医疗机构医疗服务能力基础上,为提出提升基层医疗机构检测水平的方法,探讨通过区域医疗信息化手段建立面向三级医疗机构的区域协同医疗共享平台及系列信息化系统,提升基层医疗机构检测水平,推动多级医疗协同服务模式的实现,提升区域协同医疗业务能力。     

Organizational provision for the sanitary and epidemiological safety of the population in the regions of the Stavropol Territory affected by the flood

Materials on the organizational provision and financing of measures taken with a view to ensure the sanitary and epidemiological safety in the Stavropol Territory in connection with the high flood of 2002 are presented. Due to the joint efforts of sanitary, antiepidemic and medico-prophylactic services of the Territory, administrative organs and institutions, as well as other relevant departments, not only high quality restoration works were carried out and completed in a short time, medical assistance was given to the victims and the appearance of mass infectious diseases prevented, but also the growth social tensions in the affected areas was averted.     

Quantitative motion analysis and visualization of cellular structures

The availability of cellular markers tagged with the green fluorescent protein (GFP) has recently allowed a large number of cell biological studies to be carried out in live cells, thereby addressing the dynamic organization of cellular structures. Typically, microscopes capable of video recording are used to generate time-resolved data sets. Dynamic imaging data are complex and often difficult to interpret by pure visual inspection. Therefore, specialized image processing methods for object detection, motion estimation, visualization, and quantitation are required. In this review, we discuss concepts for automated analysis of multidimensional image data from live cell microscopy and their application to the dynamics of cell nuclear subcompartments.     

基因治疗微链载体的构建及表达分析

基因载体的转染、表达效率低和存在安全问题是基因治疗的难点。由于传统病毒及质粒载体含有大量外源基因, 其表达有可能引发较严重免疫副反应。本课题旨在新的设计思路上开发高效安全基因治疗载体。 微链载体利用设计好的Cap序列封闭基因表达框两端, 起到防止细胞内核酸外切酶降解的作用。从pEGFP-N3质粒中分离纯化得到GFP基因作为微链载体的报告基因。将微链载体与原质粒载体(pEGFP-N3质粒)分别转染入真核细胞, 利用荧光显微镜和流式细胞仪检测并比较其转染效率。结果显示微链载体在293、CNE2、3T3、B95-8等真核细胞中的转染、表达效率较高, 并具有较小的细胞毒性。初步证实了微链载体在真核细胞中转染、表达效率及安全性等方面具有一定的优越性。     

Whole-cell cancer vaccination: from autologous to allogeneic tumor- and dendritic cell-based vaccines

The field of tumor vaccination is currently undergoing a shift in focus, from individualized tailor-made vaccines to more generally applicable vaccine formulations. Although primarily predicated by financial and logistic considerations, stemming from a growing awareness that clinical development for wide-scale application can only be achieved through backing from major pharmaceutical companies, these new approaches are also supported by a growing knowledge of the intricacies and minutiae of antigen presentation and effector T-cell activation. Here, the development of whole-cell tumor and dendritic cell (DC)-based vaccines from an individualized autologous set-up to a more widely applicable allogeneic approach will be discussed as reflected by translational studies carried out over the past two decades at our laboratories and clinics in the vrije universiteit medical center (VUmc) in Amsterdam, The Netherlands.     

SPM Receptor Expression and Localization in Irradiated Salivary Glands

Radiation therapy–mediated salivary gland destruction is characterized by increased inflammatory cell infiltration and fibrosis, both of which ultimately lead to salivary gland hypofunction. However, current treatments (e.g., artificial saliva and sialagogues) only promote temporary relief of symptoms. As such, developing alternative measures against radiation damage is critical for restoring salivary gland structure and function. One promising option for managing radiation therapy–mediated damage in salivary glands is by activation of specialized proresolving lipid mediator receptors due to their demonstrated role in resolution of inflammation and fibrosis in many tissues. Nonetheless, little is known about the presence and function of these receptors in healthy and/or irradiated salivary glands. Therefore, the goal of this study was to detect whether these specialized proresolving lipid mediator receptors are expressed in healthy salivary glands and, if so, if they are maintained after radiation therapy–mediated damage. Our results indicate that specialized proresolving lipid mediator receptors are heterogeneously expressed in inflammatory as well as in acinar and ductal cells within human submandibular glands and that their expression persists after radiation therapy. These findings suggest that epithelial cells as well as resident immune cells represent potential targets for modulation of resolution of inflammation and fibrosis in irradiated salivary glands.     

Virus susceptibility of Chinese hamster ovary (CHO) cells and detection of viral contaminations by adventitious agent testing

Biopharmaceuticals are of increasing importance in the treatment of a variety of diseases. A remaining concern associated with their production is the potential introduction of adventitious agents into their manufacturing process, which may compromise the pathogen safety of a product and potentially cause stock‐out situations for important medical supplies. To ensure the safety of biological therapeutics, regulatory guidance requires adventitious agent testing (AAT) of the bulk harvest. AAT is a deliberately promiscuous assay procedure which has been developed to indicate, ideally, the presence of any viral contaminant. One of the most important cell lines used in the production of biopharmaceuticals is Chinese hamster ovary (CHO) cells and while viral infections of CHO cells have occurred, a systematic screen of their virus susceptibility has never been published. We investigated the susceptibility of CHO cells to infection by 14 different viruses, including members of 12 families and representatives or the very species that were implicated in previously reported production cell infections. Based on our results, four different infection outcomes were distinguished, based on the possible combinations of the two factors (i) the induction, or not, of a cytopathic effect and (ii) the ability, or not, to replicate in CHO cells. Our results demonstrate that the current AAT is effective for the detection of viruses which are able to replicate in CHO cells. Due to the restricted virus susceptibility of CHO cells and the routine AAT of bulk harvests, our results provide re‐assurance for the very high safety margins of CHO cell‐derived biopharmaceuticals. Biotechnol. Bioeng. 2010;106: 598–607. © 2010 Wiley Periodicals, Inc.     

孕妇分娩机构选择及其影响因素分析

目的:调查孕妇分娩机构选择情况,并对影响分娩机构选择的因素进行分析,为加强分娩机构建设,合理配置助产机构医疗资源提供参考。方法:本研究调查对象均为2019年5月-2019年9月期间在我院办理建档立册的孕产妇,随机抽取420例孕妇进行自制问卷调查,实际发放问卷420份,获得有效问卷为395份。采用EpiData3.1软件建立数据库,统计孕妇基本资料、选择分娩机构考虑因素,多元Logistic回归分析影响选择分娩机构的因素。结果:90%以上的孕妇认为医疗技术水平、仪器设备水平、是否有新生儿科、医院声誉、服务态度、环境比较重要,是选择医疗机构会考虑的因素;大多数孕妇选择三级分娩机构、专科分娩机构进行分娩;经单因素和多因素分析显示,专家门诊、费用、医院级别可能是孕妇选择分娩机构类别的独立影响因素(P0.05)。结论:孕妇趋向于选择三级、专科分娩机构进行分娩,增强二级和民营助产机构的核心竞争力,有助于合理配置助产机构医疗资源,更好保障母婴安全。     

A nanoliter-scale nucleic acid processor with parallel architecture

The purification of nucleic acids from microbial and mammalian cells is a crucial step in many biological and medical applications. We have developed microfluidic chips for automated nucleic acid purification from small numbers of bacterial or mammalian cells. All processes, such as cell isolation, cell lysis, DNA or mRNA purification, and recovery, were carried out on a single microfluidic chip in nanoliter volumes without any pre- or postsample treatment. Measurable amounts of mRNA were extracted in an automated fashion from as little as a single mammalian cell and recovered from the chip. These microfluidic chips are capable of processing different samples in parallel, thereby illustrating how highly parallel microfluidic architectures can be constructed to perform integrated batch-processing functionalities for biological and medical applications.