Development of feeder-free PSC culture system enabling translational & clinical research

Pluripotent stem cell (PSC) culture using the xeno-free Essential 8™ Medium/truncated recombinant human Vitronectin system has been shown to support normal PSC properties and provide a large pool of cells for disease modeling and drug development. As research moves from translational to clinical research, general regulatory guidance from the US Food and Drug Administration (FDA) indicates that, cGMP manufactured, or clinical grade reagents should be used whenever available as ancillary reagents to minimize downstream risk to patients. Thus, we sought to identify regulatory compliant, animal-originfree alternatives for growth factors contained within the Essential 8™ Medium and incorporate ISO13485 manufacturing for the recombinantly expressed, truncated human Vitronectin (rhVTN-N), producing a qualified ancillary system for PSC expansion. Here we present data to support a seamless transition from the xeno-free Essential 8™ Medium system to the Cell Therapy Systems (CTS™) animal-origin free system. Compatibility is shown with existing cGMPmanufactured passaging reagents: Versene Solution for clumped cell passaging and CTS™ TrypLE™ Select combined with RevitaCell™ Supplement for single cell passaging. Upon expansion, PSCs are shown to maintain normal PSC properties, including morphology, pluripotency, karyotype, and trilineage differentiation potential. Together this system provides a consistent, feeder-free PSC culture medium for translational and clinical research

New viral and non-viral platforms for T-cell engineering

FDA approval of the first Chimeric Antigen Receptor T cell (CAR-T) therapy offers cancer patients more promise than ever for curative effects. However, many technical challenges in T cell gene delivery still remain in order for this therapy to become a standard of care practice. In this webinar, we will highlight the different viral and non-viral delivery approaches used in T cell engineering for cell and gene therapy applications including: New solution for small-to-large scale serum-free, suspension lentiviral production – LV-MAX™ Lentiviral Production System o Platform development process using Design of Experiment (DOE) methodologies o High-throughput to large scale bioreactor protocols o Cost benefits of this system over current methods Novel gene editing tools for primary T cells o New potent gene editing tools to increase knock-in and knock-out efficiency o Addressing non-viral delivery barriers through protocol optimization Learning Objectives: Current industry trends and challenges of cell and gene therapy manufacturing Benefits of innovative new upstream technologies for virus generation in suspensio

Superior expansion of long-term hematopoietic stem cells using StemPro™ HSC medium kit

The use of CD34+ hematopoietic stem cells (HSC) for transplantation has been limited due to the low CD34+ cell numbers in tissue sources such as peripheral blood and cord blood. Two strategies have been employed to increase the CD34+ cell dosage. These include mobilization of HSC into peripheral blood via injection of G-CSF, and ex vivo expansion of CD34+ cells. A major limitation of current systems used for the expansion of HSC is that ex vivo culture leads to expansion and differentiation of cells, at the expense of the most primitive pluripotent long-term HSC. This has limited the clinical application of ex vivo expanded HSC, since short-term progenitor cells only provide transient protection, ultimately reducing the positive health outcomes, increasing the duration of hospitalizations, and health care costs per patient. Development of a culture system that expands, both short term progenitor cells and long-term HSC would enable immune protection during the early phase of recovery, and provide a suitable solution for transfusion-independent hematopoiesis. Therefore, we have developed an HSC culture medium that enables the expansion of both long-term HSC and short-term progenitor cells, while maintaining their functional properties. We conducted several iterative rounds of Design of Experiments (DOE) involving multifactorial analysis, and mathematical modeling methods. The DOEs allowed us to identify optimal combinations and concentrations of essential media components, small molecules, and growth factors. The performance of candidate HSC expansion media were evaluated after 7 days of culture, upon which the CD34+ cells and CD34+CD90+CD45RA- cells (long-term HSC) were quantified. We were successful in developing a media system- StemPro™ HSC Medium Kit-which is xeno-free, serum-free medium that expands both long term CD34+CD90+CD45RA- HSC and short term CD34+. The expression of aldehyde dehydrogenase was conducted to identify primitive stem cells, and colony-forming unit assays were performed to assess the in vitro differentiation capacity of expanded cells. We plan to determine whether the expanded cells are engraftable by transplanting the cells into immuno-deficient mice. Taken together, we seek to highlight our design philosophy in HSC culture media development, and we believe our efforts are critical for the successful utilization of hematopoietic stem cell transplants in translational cell therapies

Implementing Preventive Chemotherapy through an Integrated National Neglected Tropical Disease Control Program in Mali

Neglected tropical diseases (NTDs) are a group of chronic infections that affect the poorest group of the populations in the world. There are currently five major NTDs targeted through mass drug treatment in the affected communities. The drug delivery can be integrated to deliver different drug packages as these NTDs often overlap in distribution. Mali is endemic with all five major NTDs. The integrated national NTD control program was implemented through the primary health care system using the community health center workers and the community drug distributors aiming at long-term sustainability. After a pilot start in three regions in 2007 without prior examples to follow on integrated mass drug administration, treatment for the five targeted NTDs was gradually scaled up and reached all endemic districts by 2009, and annual drug coverage in the targeted population has since been maintained at a high level for each of the five NTDs. Around 10 million people received one or more drug treatments each year since 2009. The country is on the way to meet the national objectives of elimination or control of these diseases. The successes and lessons learned in Mali are valuable assets to other countries looking to start similar programs

The Global Trachoma Mapping Project: Methodology of a 34-Country Population-Based Study.

PURPOSE: To complete the baseline trachoma map worldwide by conducting population-based surveys in an estimated 1238 suspected endemic districts of 34 countries. METHODS: A series of national and sub-national projects owned, managed and staffed by ministries of health, conduct house-to-house cluster random sample surveys in evaluation units, which generally correspond to "health district" size: populations of 100,000-250,000 people. In each evaluation unit, we invite all residents aged 1 year and older from h households in each of c clusters to be examined for clinical signs of trachoma, where h is the number of households that can be seen by 1 team in 1 day, and the product h × c is calculated to facilitate recruitment of 1019 children aged 1-9 years. In addition to individual-level demographic and clinical data, household-level water, sanitation and hygiene data are entered into the purpose-built LINKS application on Android smartphones, transmitted to the Cloud, and cleaned, analyzed and ministry-of-health-approved via a secure web-based portal. The main outcome measures are the evaluation unit-level prevalence of follicular trachoma in children aged 1-9 years, prevalence of trachomatous trichiasis in adults aged 15 + years, percentage of households using safe methods for disposal of human feces, and percentage of households with proximate access to water for personal hygiene purposes. RESULTS: In the first year of fieldwork, 347 field teams commenced work in 21 projects in 7 countries. CONCLUSION: With an approach that is innovative in design and scale, we aim to complete baseline mapping of trachoma throughout the world in 2015

Integration of water, sanitation, and hygiene for the prevention and control of neglected tropical diseases: a rationale for inter-sectoral collaboration.

Improvements of water, sanitation, and hygiene (WASH) infrastructure and appropriate health-seeking behavior are necessary for achieving sustained control, elimination, or eradication of many neglected tropical diseases (NTDs). Indeed, the global strategies to fight NTDs include provision of WASH, but few programs have specific WASH targets and approaches. Collaboration between disease control programs and stakeholders in WASH is a critical next step. A group of stakeholders from the NTD control, child health, and WASH sectors convened in late 2012 to discuss opportunities for, and barriers to, collaboration. The group agreed on a common vision, namely "Disease-free communities that have adequate and equitable access to water and sanitation, and that practice good hygiene." Four key areas of collaboration were identified, including (i) advocacy, policy, and communication; (ii) capacity building and training; (iii) mapping, data collection, and monitoring; and (iv) research. We discuss strategic opportunities and ways forward for enhanced collaboration between the WASH and the NTD sectors

The role of nutrition in integrated programs to control neglected tropical diseases

There are strong and direct relationships between undernutrition and the disease caused by infectious organisms, including the diverse pathogens labeled as neglected tropical diseases (NTDs). Undernutrition increases the risk of infection, the severity of disease and the risk that children will die, while the physical damage, loss of appetite, and host responses during chronic infection can contribute substantially to undernutrition. These relationships are often synergistic. This opinion article examines the role of nutrition in controlling NTDs and makes the point that mass drug treatment - the major strategy currently proposed to control several diseases - is crucial to controlling disease and transmission, but is only the start of the process of physical recovery. Without adequate energy and nutrients to repair damaged tissues or recover lost growth and development, the benefits of treatment may not be evident quickly; the effects of control programs may be not appreciated by beneficiaries; while vulnerability to reinfection and disease may not be reduced. There is substantial potential for nutritional interventions to be added to large-scale programs to deliver drug treatments and thereby contribute, within a broad strategy of public health interventions and behavior change activities, to controlling and preventing NTDs in populations, and to restoring their health

Tropical Data: Approach and Methodology as Applied to Trachoma Prevalence Surveys

PURPOSE: Population-based prevalence surveys are essential for decision-making on interventions to achieve trachoma elimination as a public health problem. This paper outlines the methodologies of Tropical Data, which supports work to undertake those surveys. METHODS: Tropical Data is a consortium of partners that supports health ministries worldwide to conduct globally standardised prevalence surveys that conform to World Health Organization recommendations. Founding principles are health ministry ownership, partnership and collaboration, and quality assurance and quality control at every step of the survey process. Support covers survey planning, survey design, training, electronic data collection and fieldwork, and data management, analysis and dissemination. Methods are adapted to meet local context and needs. Customisations, operational research and integration of other diseases into routine trachoma surveys have also been supported. RESULTS: Between 29th February 2016 and 24th April 2023, 3373 trachoma surveys across 50 countries have been supported, resulting in 10,818,502 people being examined for trachoma. CONCLUSION: This health ministry-led, standardised approach, with support from the start to the end of the survey process, has helped all trachoma elimination stakeholders to know where interventions are needed, where interventions can be stopped, and when elimination as a public health problem has been achieved. Flexibility to meet specific country contexts, adaptation to changes in global guidance and adjustments in response to user feedback have facilitated innovation in evidence-based methodologies, and supported health ministries to strive for global disease control targets