I have compiled a list of our most popular 30 Blogs for 2017. Here are the top Cell Culture Dish blogs in alphabetical order.

All About Organoids

Held this year in Boston MA, the International Society for Stem Cell Research (ISSCR) Conference 2017 brought together more than 4,000 researchers, bioethicists, clinicians, and industry professionals from all over the globe to discuss the latest discoveries and technologies in Stem Cell Research and their contributions to advancing regenerative medicine. This year, the plenary session topics included an exciting and rapidly expanding area of research—organoids. An organoid, in the most basic sense, is a miniaturized and simplified version of an organ produced in vitro in three dimensions that show realistic micro-anatomy. Unlike two dimensional (2D) cell cultures, organoids enable three-dimensional (3D) cell growth, movement and differentiation, making this technology an effective model for understanding organ development, tissue morphogenesis and the genetic or molecular basis of diseases (Xinaris et al, 2015)…

Automated Adenovirus Purity Analysis Speeds Viral-based Gene Therapy Process Development

Adenovirus-based vectors are commonly-used gene delivery vehicles, especially in Gene Therapy applications where their efficient nuclear entry mechanism and low pathogenicity for humans are much-valued attributes. In addition, Adenovirus vectors can be produced in high titers and the particles are stable yet easy to modify.

Several methods of manufacturing Adenovirus-based products are in use today, including small-scale production in cell culture flasks, suspension-based scalable platforms and large-scale batch manufacturing in fixed-bed bioreactors. Downstream purification steps include chromatography and filtration, the objective of which is to produce high-quality, high-purity Adenovirus particles in a scalable manner. Methods for analyzing the purity and integrity of the Adenovirus particles in a time and cost-efficient way are therefore critical for this task…

Automated Collection and Analysis of Bioreactor Samples to Enable Quality by Design Initiatives

For QbD to work effectively you must accurately measure and be able to maintain tight control of your bioprocess conditions through input of reliable analytics, characterization of influencing factors, minimizing user interference and early detection of deviation. As a result, innovation in these areas has expanded what is possible in terms of bioprocess monitoring. There are two important tools required for automated culture monitoring, the first is having an automated, scalable and sterile way to collect samples from bioreactors for culture monitoring and the second is being able to analyze these samples to provide routine updates on how the culture is performing. By monitoring cell culture performance in real-time, you can not only gather information about the process, but also utilize this information to make changes to the culture as needed…

Automated Optimization of IgG Production in CHO Cells

An ideal approach to media optimization is using a factorial design of experiment (DOE), where a variety of media components are tested at different concentrations in combination with one another. However, these factorial experiments rapidly increase the number of conditions that require testing. Common ways of quantifying the production of IgG or other proteins are frequently labor intensive (i.e. ELISAs) or prohibitively slow (i.e. HPLC), particularly at the high throughputs required for DOE. In contrast…

Cell Culture Basics – Mycoplasma 101 – A practical guide to prevention, detection and elimination of mycoplasma contamination

Mycoplasma contamination is a very real concern for research labs. Mycoplasma contamination can compromise cell culture and experiment results, thus wasting valuable resources conducting experiments that ultimately aren’t accurate. Because mycoplasma contamination actually affects the cells’ overall behavior, contamination can result in false interpretation of experimental results and undermines the validity of the resulting data…

Cell Culture Monitoring – A Critical Component for Quality by Design in Cell Therapy

Most biopharmaceutical companies have implemented Quality by Design (QbD) and Process Analytical Technology (PAT) initiatives to address similar challenges in CHO-based biologics manufacturing. A QbD approach provides a systematic look at process development, which attempts to predict unique challenges and address them in advance. A key aspect of QbD is defining Critical Quality Attributes (CQA) and Critical Process Parameters (CPP) and then monitoring these closely through the use of PAT. The FDA has stressed the importance of their Quality by Design and Process Analytical Technology initiatives in an effort to encourage more consistent manufacturing processes and product quality.

While the path for implementing this approach in Cell Therapy manufacturing is arguably more complex, it would undoubtedly improve the process. However, for QbD to work effectively you must accurately measure and be able to maintain tight control of your culture conditions through input of reliable analytics, characterization of influencing factors, minimizing user interference and early detection of deviation. A critical first step of applying this approach in Cell Therapy is to ensure that the culture can be monitored precisely, so that critical process parameters can be accurately measured and ultimately maintained during manufacturing…

Continuous Processing Optimization with Smarter Tools

Due to a paradigm shift in the pharmaceutical industry, there is rising pressure to come up with faster, more cost- effective ways to produce drugs for the patients who need them. As orphan drugs and personalized medicine begin to replace traditional blockbuster products, pharmaceutical companies are looking at new and innovative ways to quickly and efficiently deliver drugs to target populations in the thousands rather than the millions. In addition, the need for lower drug prices has been pushed into the spotlight not just by regulators, but also by the advent of biosimilars. As a result of these changes, industry experts must find  a way to produce drugs that address the issues around both drug pricing and time to market while also maintaining quality and profits…

Continuous suspension cell culture monitoring in bioreactors using quantitative phase imaging

Cell culture monitoring for cell count and cell viability typically involves manual sampling from each bioreactor followed by Trypan-blue cell exclusion. This sampling needs to happen at least once per day and ideally more often. An operator must then enter the results into a spreadsheet or other tracking software and generate a growth curve. The challenge with this process is that it is highly manual, and time consuming. Sampling an entire facility means a whole team is required to monitor what can be upwards of 50+ bioreactors. In addition, manual sampling creates an opportunity for contamination and variability…

Development of a Chemically-defined, Animal Component-free Medium for T cell Culture

As an industry, biopharmaceutical manufacturing has moved away from using animal components in media formulation. This move was largely motivated by the desire for increased consistency, an improved safety profile, and a simplified regulatory pathway. Gene and immunotherapies are now increasingly making their way to clinical trials and once again, there is a desire to move away from media that contain animal-based and other undefined components. Even more important in these therapies is the impact that animal components and their naturally occurring cytokines and growth factors can have on cells…

Development of Animal-free Peptones for Mammalian and Microbial Culture

Cell culture media optimization continues to play a critical role in both increasing titers and providing a mechanism for modifying protein quality. One way to optimize media is to include supplements that improve productivity or protein characteristics. As a result, there is an interest in developing novel animal-free supplements that can impact these processes for both mammalian and microbial culture. It is important to note that supplements chosen to enhance media can have dramatic effects, both positively and negatively, on productivity, growth and protein quality. This highlights the importance of understanding what the drivers of the cell culture process are in order to identify appropriate supplements…

Development of DoE based fed-batch strategies for high-producing CHO cell cultures

There are a number of tools that are used to inform decisions about feed strategy. One of the most beneficial is to utilize Design of Experiments (DoE) to test the performance of media with various feed supplements at different concentrations. A good DoE can help determine not only which feeds are best, but also concentration and timing for adding feed components.

Many current platform processes feed a fixed-volume bolus of concentrated supplements at scheduled intervals. However, recent developments show that there may be more effective methods for scheduling feeding. A more dynamic feed strategy that responds to the needs of the culture rather than feeding on a set static schedule could yield better results…

Development of an Improved Poloxamer 188 – Optimized for Cell Culture Performance

Poloxamer 188 is a surface-active non-ionic polymer used in cell culture media as shear protectant. With time it became a standard ingredient in cell culture media for commercial production processes. It was demonstrated to increase the robustness of mammalian cells to shear from sparging, which probably is the strongest contributor to the hydrodynamic stress in bioreactors.

As mammalian cell culture technology improved, e.g., with process intensification through increasing cell densities and productivities in fed-batch and perfusion, issues with Poloxamer started to be reported. Several biopharmaceutical manufacturers reported unexpected loss of cell density and viability in their manufacturing operations which they could correlate to lot-to-lot variation in Poloxamer 188.

This variation became a major pain point in the biopharmaceutical industry and extensive investigations have been initiated to understand the source of lot-to-lot inconsistencies of Poloxamer 188.

The development and validation of proprietary analytical and biological methods based on a reference library of 100+ blinded customer and supplier samples has created the expertise to identify the critical properties of Poloxamer 188…

Direct Inoculum of Bioreactors with CHO Cells from Frozen Seed Bags to Eliminate Continual Seed Trains and improve Facility Utilization

Flexibility in cell culture manufacturing via a reduction in process duration can be a key strategy for maximizing facility utilization and facilitating the production of multiple therapeutics from a facility. A key bottleneck is the seed train, which can add weeks to the timeline of the production culture. Seeding production bioreactors with a direct, cryopreserved CHO cell inoculum could possibly eliminate the need for a lengthy, continuous seed train and provide other numerous benefits…

Efficient Large-Scale Expansion of MSCs for Translational Medicine and Research Use

Successful expansion of MSCs is important due to the popularity of the cell type in Cell Therapy applications. There are over 700 current or concluded clinical trials using MSCs worldwide, with 150 trials taking place in the United States and Canada.

Historically FBS has been used to culture MSCs, but as these cells are increasingly being used in Cell Therapy applications, there has been an interest in finding animal-free supplement alternatives. However, when these cells are used in therapeutic applications, quality and safety are a top priority. In addition, they must be expanded to cell numbers that can meet the therapeutic cell dosage levels. This requires that media and supplements must meet high quality, safety and performance standards…

Fast tracking a QbD Approach

QbD starts with an understanding of the quality target product profile. This allows potential critical quality attributes (CQAs) to be established and a risk assessment performed that links material attributes and process parameters to CQAs. By understanding the impact of process parameters on CQAs a design space can be developed in which CQAs will be met. Bioprocess engineers must then implement a suitable control strategy that ensure the process operates with the design space. Continuous improvement activities can be performed throughout the product lifecycle and enhance product quality still further…

Fed-batch culture – Optimizing feed strategies now and in the future

Great strides have been made in fed-batch culture and feed strategies with new tools and strategies introduced regularly. In addition to improving cell growth and viability, our optimization focus has grown to include strategies for influencing protein quality, such as glycosylation profiles. This article and accompanying interview will explore current feed design strategies and will look ahead to the future of fed-batch culture…

First In-Human Allogeneic Clinical Trial Commences with iPSC-derived Mesenchymal Stem Cells

It is widely accepted that stem cells can be divided broadly into embryonic and non-embryonic stem cells. Embryonic stem cells (ESCs) are derived from the inner cell mass of blastocysts and are pluripotent, meaning they can differentiate into cells of all three germ layers: ectoderm (outer layer), mesoderm (middle layer), and endoderm (inner layer). Conversely, non-embryonic stem cells are found in the extra-embryonic tissues (placenta, umbilical cord blood and amniotic fluid) and in all adult tissues, (i.e. bone marrow, fat, kidney, etc). Human mesenchymal stem cells (hMSCs) are an example of non-embryonic stem cells and were first isolated in the bone marrow and characterized by Friedenstein and his colleagues in 1974 (Amorin, 2014). hMSCs, also called mesenchymal stromal cells, are a subset of non-hematopoietic adult stem cells that originate from the mesoderm (Kim et al, 2013). They are considered to be multipotent; able to self-renew and generate progeny of several distinct cell types. Specifically, they produce cells found in the connective tissues such as cartilage cells (chondrocytes), bone cells (osteoblasts) and fat cells (adipocytes)…

Glycosylation Overview and How to Control Glycosylation using In Vitro Glycoengineering

Glycosylation is the most common type of post translational protein modification. It is also an important factor in manufacturing therapeutic proteins, depending on the mode of action, because it can impact protein stability, biological activity, and the pharmacokinetics of the final product. Due to the impact that glycosylation can have on the therapeutic, it is frequently identified as a critical quality attribute. Thus, it is important to control glycosylation during manufacturing to ensure consistent glycosylation for the final product.

GMP Proteins for Cell Therapy Manufacturing: Top 6 Things to Know

Using cells as a therapeutic offers one of the most promising new approaches for treating human disease. These methods include stem cell therapies and regenerative medicine, as well as immune cell therapies for cancers and immune-related disorders. Despite the promise, it has been difficult to bring these therapies mainstream. They are vastly more complex than traditional small molecule treatments or biotherapeutics. For example, in autologous therapies like CAR T, cells are removed from the patient, enriched for the cell of interest, genetically engineered to target the tumor, expanded in cell culture, and re-introduced into the patient as the therapeutic (Figure 1). Many of these steps have yet to be standardized and, along with the inherent variability in biological systems, there is increased potential for unanticipated results and risk to patients…

The Importance of Resin Selection in Development of a Platform Bioprocess Film

In developing single-use films that are specifically designed for biomanufacturing, the material used for construction is key and requires a great deal of research and testing. Single-use containers are constructed from plastic films, which are often composed of several layers of polymers with additives for processing and performance. The flexural properties of these layers are important to performance.

In addition, the diversity of applications for single-use containers requires film that can achieve a wide variety of performance attributes such as mechanical strength, flexibility, biocompatibility, and suitable gas barrier properties to name a few. Thus, the right balance of chemical composition and film architecture is critical for achieving desired performance across many applications…

Laminin cell culture matrices – The key to efficient derivation and reliable culture of stem cells and specialized cells lies within these extracellular matrix proteins

Reliable derivation and expansion of high quality hPSCs under feeder-free, defined and xeno-free conditions, rapid scale-up, efficient and reproducible differentiation and maintenance – all these areas are important for a Stem Cell Researcher, but many struggle to achieve it. Furthermore, for therapeutic applications, all these steps must comply with clinical requirements. Mimicking the natural cell environment is the key for success and the culture substrates play a vital role. Here, I will discuss the use of biologically relevant, human, recombinant laminin cell culture matrices from BioLamina, and how they can be used to recreate the cell-specific, physiological microenvironment on a cell culture dish. We call it biorelevance. Let me show you how laminins can help pave the way towards successful Cell Therapy…

Levers to Increase Antibody Titer While Also Improving Glycosylation

There are multiple factors that drive glycosylation and several currently available approaches to influence glycosylation patterns. Notable factors that influence glycosylation include the following: cell culture process conditions, ingredients in cell culture, genetic selection or engineering, and directly impacting the physiological organelles responsible for glycosylation (endoplasmic reticulum – the ER – and Golgi apparatus). In cell line development there are opportunities to use genetic engineering tools to manipulate the glycosylation pathway. In upstream development, cell culture process conditions (ie. temperature and pH) can be manipulated. Lastly, commercially available constituents in the glycan process, such as precursors and enzymes, can be added to cell culture media to further hone glycoprofiles. However, there have been limited available approaches to improve the functionality of the ER and Golgi…

A Look at the Current State of Continuous Bioprocessing

Recently, BioProcess International in conjunction with Pall Life Sciences published an informative supplement titled, “Is Continuous Bioprocessing the Future? ”. The publication contains 12 articles covering various aspects of continuous bioprocessing as well as additional articles on enabling technologies. I found the supplement to be a useful update on continuous bioprocessing technology benefits, implementation challenges and successes, as well as what the future holds. Following is a sampling of articles with key highlights…

A New Chemically-defined Medium for use with Suspension-adapted Avian Cells in Vaccine Manufacturing

The EB66 avian cell line, developed by Valneva, Inc. from duck embryonic stem cells, has been investigated for use in virus production with very exciting results (Madeline et al 2015, Léon et al. 2016). EB66 cells can be grown in serum-free suspension culture at high cell density, enabling much easier and more efficient scale-up than with attachment cells. More importantly, virus production levels using these cells have been comparable to or better than those obtained using attachment cells.

Interest in exploring new cell lines for vaccine manufacturing, in particular the EB66 cell line, has been increasing. In June 2016, Valneva announced a new R&D collaboration with GlaxoSmithKline (GSK) for pre-pandemic influenza vaccine based on the EB66 cell line. Prior to that, the first human vaccine using the EB66 technology received marketing approval in 2014, with the first veterinary vaccine receiving approval in 2012…

A Novel Approach for Expansion of High Quality Mesenchymal Stem Cells

Human mesenchymal stem cells (hMSCs) are a promising tool for therapeutic applications in cell-based therapy and regenerative medicine. Increasing evidence has shown that MSCs represent new hope for treating, and in some cases, perhaps even curing human diseases like diabetes, Alzheimer’s, Parkinson’s, Muscular Dystrophy , inflammatory disorders, osteoarthritis and damaged cartilage, to name a few.   And although stem cells have emerged as the future of health care there are still a number of unmet needs, which must be addressed before widespread clinical and therapeutic applications become a reality. One of the major challenges in Cell Therapy is obtaining sufficient numbers of quality stem cells while maintaining their differentiation potential. BM-HPME^® (Bone Marrow – High Performance Micro Environment) overcomes key obstacles to capturing and growing stem cells without losing potency…

A Novel 3D Immune Oncology Model for High Throughput Testing of Tumoricidal Capability

Immunotherapy, particularly for cancer treatment, is a very active area of both academic and clinical research. There has been increased interest in utilizing a patient’s own T cells to attack their tumors. However, positive laboratory results do not always translate from bench to the clinic, and the effectiveness of immune cells therapies are not equivalent for all patients or cancer types. To better understand these differences, there is a need for improved in vitro oncological models that more closely resemble the tumor environment in vivo. Traditionally, tumoricidal activity and immune evasion have been studied by utilizing two dimensional systems (2D) which may not accurately reflect the complexity of a three dimensional (3D) tumor. The physical barriers immune cells need to overcome in a 3D system are much greater than those of 2D. Not only do immune cells need to migrate to the tumor site, but they also need to infiltrate a 3D structure in order to attack the target cells. Furthermore, it has been shown that phenotypic differences occur in 3D-cultured tumor cells that allow for a higher resistance to cytotoxicity^1-2. Here, we demonstrate a novel, high-throughput 3D model to study cancer/immune cell interactions by combining two commercially available products; 96 well ultra-low attachment microplates and 96 well permeable support systems. By replacing the standard 2D flat-bottom permeable support receiver plate with an ultra-low attachment microplate, an easy-to-use, 3D, high-throughput assay to investigate immune cell homing, tumor cytotoxicity, and tumor immune evasion is achieved…

Reproducibility and Antibody Articless in Research – Addressing the reproducibility crisis

Reproducibility is a pillar of research. Studies must be reproducible to have merit. This is for good reason, as research is foundational, one study builds upon another and so on with each group contributing their piece to the overall puzzle. At least this is the way it is supposed to work. However, when research is reported erroneously, it creates a tremendous waste of time and resources for the entire research community working on the same application. While concerns about reproducibility have been discussed for years, the problem more recently has escalated to what is frequently called a “reproducibility crisis”.

Synergizing Transient and Stable Protein Expression for Accelerated Biotherapeutic Development

Early in development, small amounts of protein are needed to conduct preliminary studies that inform a go/no go decision. Protein can either be generated using transient production or through stable cell line generation. Most often companies choose to use transient transfection in either HEK or CHO cells to produce small quantities of protein because it is faster and more cost effective than developing a cell line. Having access to preclinical material faster and with less cost can greatly impact the overall drug development timeline. Then if the product progresses further in development, a stable cell line will be generated for larger scale manufacturing.

However, the timing of when to generate the stable cell line can be tricky. Stable cell lines are labor intensive and costly to develop. If companies develop one too early, the product may still fail preliminary studies and the time and money would be wasted. If companies develop the stable cell line too late, they may run into manufacturing, quality and timing delays…

Transcriptome analysis reveals strategies for CHO cell culture media design and feed-spiking strategy to improve batch culture

When looking at batch vs. fed-batch culture, there are advantages and disadvantages to each, so considering the application is very important. Batch cultures are fairly simple, straightforward and take very little time to set up. However, batch culture typically doesn’t yield high mAb titers, due to nutrient depletion, by-product accumulation and short growth and production phases. So a batch approach may be good for lab-scale processes where product needs to be generated quickly and simply with little optimization. In contrast, with fed-batch culture, feeds are added to replenish nutrients, which increases cell concentrations, process time, and can yield much higher titers. However, fed batch culture also requires more time for optimization and resources to run. So this approach is more beneficial in situations where you would are developing long term processes or scaling up to large-scale production…

Video – Upstream Bioprocessing Cell Culture Overview

An overview of upstream bioprocessing steps beginning with an explanation of molecular biology and cell line engineering, then continuing with creation of a working cell bank, cell expansion, bioreactor production and clarification steps.

Coming soon – Top Cool Tool Features

Don’t miss our Top 25 Downstream Column Blogs of 2017