I have compiled a list of our most popular 30 Blogs for 2016. Here are the top Cell Culture Dish blogs in alphabetical order.

3D Bioprinting and 4D Bioprinting – Applications and Technologies

3D bioprinting, a type of additive manufacturing, is the process of creating a three-dimensional (3D) structure by laying down successive sheets of living cells that build upon each other. The specific mechanical means of this deposition of cells and matrix vary greatly between bioprinters and bioprinting applications. The printing material used in bioprinting is often called “bio ink” and consists of living cells and material needed to support cell growth and proper tissue structure. The exact bio ink formulation varies depending on the cells being used and the structure being created. The next generation of 3D bioprinting is called 4D bioprinting…

Another Perspective on 4D Bioprinting

3D bioprinting has been promoted and even implemented to varying degrees for more than a decade [1]. Proposed applications abound, although for some their component technologies remain to be completely established [2]. The recent invention of 4D printing naturally points to the general concept of 4D bioprinting. However, the principles and terms inherent to the concept of 4D bioprinting are still in early development. Here an outline of the proposed implementations of 4D bioprinting is reviewed and a comprehensive definition introduced…

Antibody Drug Conjugates – An Overview of the Technology and Manufacturing

Antibody drug conjugates are composed of three parts:an antibody specific for the tumor associated antigen, which has restricted expression on normal cells, a cytotoxic agent designed to kill target cells when internalized and released and, a chemical linker to attach the cytotoxic agent to the antibody. New linker systems are designed to be stable in circulation and release the cytotoxic agent after being internalized by the targeted cells. Because the cytotoxic agent is delivered in such small quantity, it must be powerful enough to kill the tumor cells. Therefore, the potency of these cytotoxic agents is 100-1000 fold more potent than cytotoxic agents delivered as free drugs. The real benefit is the ability to deliver a very potent cytotoxic agent only to the targeted tumor cells…

Cell Culture and Single Cell Passaging of Human Pluripotent Stem Cells Without the Need for ROCK Inhibitor

Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs), and human induced pluripotent stem cells (hiPSCs) have the ability to self-renew and to give rise to specialized cell types and therefore, have tremendous potential in clinical, drug discovery, and regenerative medicine applications¹. Human PSCs are traditionally cultured on a layer of feeder cells (such as mouse or human fibroblasts)². For feeder-free cultures, these cells are cultured on either complex mixtures of naturally- derived extracellular matrices (such as Corning Matrigel® matrix), or more recently on a variety of recombinant proteins (such as Laminin) and synthetic substrates (such as Corning Synthemax® II-SC)²…

Cell Culture Basics: Stem Cell Media – The “What” and “Why”

For over 30 years, Biological Industries has been developing specialized cell culture media for many cell types, including primary cells and stem cells. Over time we have seen a significant evolution of stem cell media as well as our ability to understand, evaluate, and optimize cell culture conditions. Cell culture is one of the most common and complex techniques used in the life sciences, and the media itself is a critical element for maintaining healthy, proliferating stem cells in culture. When you break it all down, all stem cell media essentially contain the same basic components: a basal medium, buffer system, glutamine, serum (or serum alternative), specific growth factors, and additional supplements. Here we look at a few of the main media components to better understand their importance and influence on cell cultures and how working with a highly pure and defined stem cell medium is key for regenerative medicine applications…

Cell Flask Adapters Can Streamline the Cell Culturing Process

During various stages of the cell culturing process, centrifugation is frequently used to isolate extracellular products or to separate cells from their aqueous environment. Traditionally, cells and media are first transferred from a cell culture flask to a 15 mL or 50 mL conical tube before centrifugation. These transfer steps require operator labor and time, as well as the cost of the transfer vessel, and also introduce the potential for contamination during transfer. With cell culture flask adapters, however, the culture can be centrifuged directly in the flask. This helps streamline the cell culturing process by decreasing time and labor. Moreover, it reduces the potential for contamination and decreases overall labware costs…

Cell Therapy for Parkinson’s Disease – An update on the move toward clinical trials

This past June Cell Culture Dish attended the International Society for Stem Cell Research (ISSCR) Annual Meeting in San Francisco and over the next several weeks we will be sharing information that we gathered. One topic that I was happy to see covered was an update on the clinical progress of a Cell Therapy for Parkinson’s Disease. When I attended ISSCR in 2014, I had listened to an interesting talk by Dr. Roger Barker, John Van Geest Centre for Brain Repair and Department of Neurology, University of Cambridge, titled “Taking Stem Cell-Based Therapies to the Clinic in Parkinson’s Disease”. I followed up the talk with a blog titled, “Cell Therapy for Parkinson’s Disease – Considerations for the Future,” that discussed the status of Cell Therapy opportunities at that time…

CHO Media Development and Optimization – A look at three case studies

CHO media development and optimization is always a popular topic since media plays a key role in biomanufacturing success. In my BPI East coverage last month, “Biomanufacturing CHO Media – A look at different approaches and optimization opportunities,” I discussed different media development and optimization strategy options, highlighting decisions that companies need to make when thinking about how they will handle their CHO media development from scale up through commercialization of product candidates. These decisions must take into account many factors including, yield, product quality, timelines, impact on downstream purification, regulatory compliance and raw material sourcing, just to name a few. Companies may choose to develop their own platform media for their products, use an off the shelf media, partner to create a custom formulation or optimization, or take a hybrid approach…

The CRISPR/Cas9 System and its Applications

Genome editing in vitro has been studied since the 1970’s with the discovery that exogenous DNA could be taken up by yeast or bacteria and randomly integrated into the genome. Subsequently, targeted integration of DNA into the yeast (Saccharomyces cerevisiae) genome was demonstrated (Scherer et al., 1979), opening the doors for genetic manipulation. Since then, progress in applications of genome editing (i.e. Gene Therapy) has been limited due to lack of efficient, inexpensive, and accurate methodologies…

CRISPR Technology Where it Stands and What the Future Holds

Previously, I wrote an article summarizing the CRISPR/Cas9 system (CRISPR short for Clustered Regularly Interspaced Short Palindromic Repeats) and its applications. This prokaryotic defense mechanism (first discovered in 1987) which cleaves the DNA of invading phages and plasmids has revolutionized how we perform genetic modifications in eukaryotic cells since it was first published in 2013. It has enabled researchers to use the Cas9 ‘molecular scissors’ to cut DNA at a specific locus as directed by a guide RNA sequence. The technology has been rapidly accepted by researchers worldwide. For example, at the International Society for Stem Cell Research (ISSCR) conference this year in San Francisco CA, 90 of the 504 posters presented mentioned the CRISPR/Cas9 technology. That’s 18% of all the poster presentations spanning many different cell types and disciplines. It is amazing to see such a quick adoption of a technique in just three short years but certainly not surprising as researchers continue to be excited about the potential of CRISPR-based gene editing. It is a game-changer in the molecular toolbox, enabling efficient, cost-effective, precision gene editing that was simply not attainable before…

Digital Biomanufacturing Will Enable Tissue Bioprinting

The digital revolution in manufacturing began with the recent explosion in process monitoring capabilities. This generated immense amounts of data about increasingly greater aspects of the biomanufacturing process. PAT-driven innovations occurring here now include new fluorescent and IR based probes and automated cell-free sampling devices that can direct samples to any number of advanced analytics in real time. Combined with such new information gathering and interfacing techniques from advanced computer hardware, OPC and cloud capabilities– we are seeing a qualitative change in our data gathering and sorting capabilities…

Electroporation-based Transfection Demonstrates Consistent Antibody Quality and Glycosylation Patterns for Biotherapeutic Product Development

Increasingly, companies are choosing CHO-based transient production of antibodies in early biopharmaceutical development over stable cell line generation. The advantage of using transient transfection is that it takes significantly less time and cost to generate material when compared with the alternative of developing a genetically stable cell line for manufacturing. This is particularly important in drug development where preclinical material is needed quickly in order to make informed go/no go decisions. Having access to preclinical material faster and with less cost can greatly impact the overall drug development timeline…

Enabling High Density Cell Banking using a Single-Use, Closed-System

Working cell banks are typically the first step in any biotherapeutic manufacturing campaign. Thus their efficiency is key in getting product manufactured in the shortest timeline possible. In recent years the industry has turned attention toward increasing the efficiency of cell banking, inoculation and seed train scale up. Key areas identified as areas for improvement include: high density cell banking with banking volumes up to 1 liter and closed systems to reduce the risk of contamination and manual operations…

Generating Recombinant Versions of Human Serum-Derived Proteins – Transferrin and Albumin

Albumin in Cell Culture Media – An examination of quality and function

Albumin Fatty Acid Profiles for cell culture media – Enabling Albumin Optimization for Cell Culture Media

Three part recombinant serum derived protein series – Due to the multifaceted biological role of human serum albumin, inclusion in cell culture media has been shown to be extremely beneficial for the propagation of many cell types ex vivo. However, proteins isolated from human serum can have variable performance and potential adventitious agent contamination risks. Thus, the investigation into the inclusion of a recombinant alternative in serum free cell culture media is warranted. Due to the inherent complexity of albumin function, the successful substitution of human serum-derived albumin with recombinant versions in cell culture media can be somewhat involved. One particular function, fatty acid binding and subsequent delivery to proliferating cells, can have extremely profound effects in cell culture and is worthy of further discussion…

Gene Therapy Strengthened by Recent Successes

April has been a busy month for Gene Therapy news, the most significant being the decision of a European Regulatory Committee to recommend approval of GlaxoSmithKline’s Gene Therapy for the treatment of “bubble baby disease”. If approved, this would be the first Gene Therapy approved for children. While GSK’s therapy certainly appears to be the most likely next approved Gene Therapy, there are many others making their way through clinical trials. According to a recent article published by Regulatory Focus, “FDA Sees Spike in Gene and Cell Therapy Applications,” the number of gene and cell therapies in clinical trials is growing and FDA is utilizing their Cellular, Tissue and Gene Therapies Advisory Committee (CTGTAC) to address concerns about handling the increase in applications…

Increase Protein Yield by Striking the Right Balance between Cell Density and Protein Productivity

How do proliferation rate and VCD relate to protein titer, if at all? In this blog, we examine the relationship between cell growth and protein productivity with the help of a study evaluating cell metabolism in fed-batch CHO cell culture…

New Advances Pose New Challenges to Bioproduction

The sales of biologic drugs have surpassed $125 billion. As the market has grown, increased competition between companies making therapeutics for the same target, such as TNF-α, is occurring. Additional price pressure has been brought on by the entry of biosimilars, furthering the need to reduce manufacturing costs. One way to reduce costs is to increase manufacturing productivity. Many tools exist to increase productivity, and in each case media optimization is required as part of the program. Often scientists focus on maximizing protein production, but there are potential consequences for focusing only on increasing protein titer. In this article, we discuss some of the available tools for increasing productivity, the complexities associated with reducing biomanufacturing expenses and potential solutions that are available to address these issues…

New Poster: Proving Clonality – A Documented Clonality Report for Regulatory Submission

As part of our ongoing poster coverage, we will be writing about some of the posters presented at various conferences. Solentim recently presented a poster at Cell Culture Egineering XV that I thought provided a great solution for documenting clonality to the regulatory agencies. The poster entitled “Proof that can travel – Documented Clonality Report for Regulatory Submission” discusses Solentim’s solution to documenting clonality for regulatory submissions in the form of an easy, rapid report…

Particulates in Cell Therapy Products – An important issue for commercialization

The presence of particulates in Cell Therapy products is an issue that is becoming increasingly important as potential cell therapies move through the clinical pipeline. I recently read a very informative paper on this topic titled, “Managing Particulates in Cell Therapy: Guidance for Best Practice,” published in Cytotherapy and written by the International Society for Cellular Therapy (ISCT) Process and Product Development Subcommittee (Clarke, et al.). The paper was written to provide guidance for Cell Therapy developers, manufacturers and also suppliers regarding the management of particulates in Cell Therapy products. This paper is particularly valuable, since currently there are no set limits or standards specific to particulates and Cell Therapy products…

Poster: Convert your glass benchtop bioreactors to single-use with minimal capital investment

As part of our BPI West 2016 coverage, we will be writing about some of the posters presented at the conference. One poster that I thought provided a great way to save on capital expenditure and give existing equipment an update and new life was presented by Distek, “The BIOne benchtop single-use bioreactor system for mammalian cell growth and recombinant protein production provides a robust model for bioprocess development.” In the poster, Distek describes their single use benchtop bioreactor system that allows users to convert existing glass benchtop bioreactors to single use bioreactors with minimal capital investment…

Poster: Flow Electroporation Provides a Highly Efficient, Flexible and Scalable Transient Protein Expression System

Transient transfection has been used to quickly and cost-effectively manufacture milligram quantities of recombinant protein in HEK 293 for years. The advantage of using transient transfection is that it takes significantly less time to generate material when compared with the alternative of developing a genetically stable cell line for manufacturing. This is particularly important in drug development where preclinical material is needed quickly in order to make informed go/no go decisions. Having access to preclinical material faster can greatly impact the overall drug development timeline.  The challenge has been the transfection specifically of CHO cells to express the proteins of interest. CHO cells are typically more difficult to transfect, but are the preferred vehicle of antibody production due to their use in downstream production of clinical material…

Predicting Differentiation and Characterizing Pluripotent Stem Cells Using Non-invasive Multi-analyte Luminex® Assays

One of the challenges facing Stem Cell Research and Cell Therapy applications today is characterizing and investigation of stem cell and differentiated populations. There are several options currently available for this type of investigation, for example, rt-PCR, western blot, immunochemistry, and flow cytometry. However all these methods are invasive, which means that valuable cells are lost during the process of characterization…

Proving Clonality is a Hot Topic at Informa “Cell Line Development and Engineering” Conference

The meeting opened with a talk by the predominant industry regulator, the US FDA, and ended with group workshop, chaired by former FDA CMC reviewer Dr. Audrey Jia, evaluating and providing feedback to the wider audience on the various strategies used by industry to demonstrate monoclonality in their filings. The discussion clearly indicated that within the cell line development arena there is truly a mixed bag of methods being employed for tackling the clonal production of cell lines ranging from the traditional multiple rounds of limiting dilution with statistical analysis, to the growing trend of reducing cloning rounds via the introduction of automated cell seeders and imaging solutions…

Pumping Iron – But Not in the gym: The Critical Roles of Transferrin in Cell Culture Media

Since the early days of cell culture, scientists have typically relied on serum supplementation in their cell culture media, most often in the form of whole bovine or human-derived serum or isolated components/fractions thereof. This ill-defined mixture of proteins, small molecules, and other factors has served as a physiological crutch compensating for our limited understanding of the complex ingredient interactions required for in vitro propagation of mammalian cells. As cell culture-based therapeutics have transitioned from bench research to clinical trials at bed side, they have established the desperate need for a great “cell cultural enlightenment” – and our days of blissful ignorance must come to an end…

Reduce Cell Line Development Time by 30% and Simplify Proof of Clonality – A Case Study

Cell line development for biomanufacturing is time consuming and typically the rate-limiting step is filing and IND submissions. Demonstrating clonality of the cell line is a critical part of the overall regulatory package. Even though 2 rounds of limiting dilution have been deemed acceptable in the past, new technologies have enabled proof of clonality to be demonstrated through captured images of the cells at various stages of the process, thus eliminating the need for a second round of limiting dilution. By eliminating the second round of limiting dilution, companies are able to significantly reduce their timelines, whilst meeting regulatory requirements early in the new drug development process. Biotherapeutics company, SystImmune, presented their innovative approach to Cell Line Development at the Cell Line Development & Engineering Conference held by IBC in San Francisco in June…

Solve Production Challenges of Difficult to Express Proteins with Scalable, Continuous Manufacturing

At this year’s Boston Biotech Week there were many interesting talks on continuous biomanufacturing and associated new technologies. In particular, there was a great deal of discussion around how to handle difficult to produce proteins. These manufacturing challenges included proteins that were difficult to express and/or were unstable…

Ten Years of Induced Pluripotent Stem Cells: A Look Back

This year, I had the opportunity to attend the International Society for Stem Cell Research (ISSCR) conference held in San Francisco, CA. It was wonderful to hear about the ‘hot topics’ in Stem Cell Research and to see new technologies from industry leaders. On the second day of ISSCR2016, I had the pleasure of hearing Shinya Yamanaka give a talk on induced pluripotent stem cells (iPSCs). The title of his talk was ‘Reprogramming of Cells and Scientist’. The first part was clearly autobiographical while the second half was focused on basic stem cell science (NAT1’s role in pluripotency). It was clear he was reflecting on how iPSC technology has changed him as a scientist as well as the scientific community as a whole. This year marks the ten year anniversary of the first induced pluripotent stem cell paper by Shinya Yamanaka and Kazutoshi Takahashi. He was awarded the 2012 Nobel Prize in medicine, which he shared with John B. Gurdon, for this revolutionary discovery…

Upstream Continuous Process – Perfusion Culture

Continuous processing has proved a very successful model in many other industries. As such, there has been a growing interest in utilizing continuous process concepts in the manufacture of biopharmaceuticals. Furthermore, certain continuous technologies have already been incorporated into existing biopharmaceutical manufacturing with many benefits. For example, in several applications, perfusion culture has provided benefits over traditional fed-batch processing and has been used successfully for many years to manufacture unstable proteins. These benefits can include higher yield, increased speed, cost savings, efficient facility utilization, better scalability and improved product quality and stability…

The Use of Animal Serum in the Clinical Translation of hMSCs

Human mesenchymal stem or stromal cells (hMSCs) are an integral part of cell-based therapeutics, with over 400 clinical trials recently completed or in progress using hMSCs. As more research teams transition their stem cell-based regenerative technologies to the clinic, the use of serum in the cell production process has been, and will continue to be, a necessary evil that must be managed.  Luckily, pharmaceutical regulatory agencies, driven by the biologics industry over the last 30 years, have established guidances and guidelines that have helped to demystify and clarify some critical aspects of dealing with animal components. As it is important to have an understanding of how to manage serum during the clinical translation of hMSCs, we have focused this blog post on this specific topic…

What does Xeno-Free really mean, and why does it matter to cell culture scientists today?

Despite the fact that xeno-free is a term most cell culture scientists hear regularly, there is considerable confusion around what that term actually means. Ever since the dawn of the effort to remove fetal bovine serum from media due to its complex and variable nature, there has been a variety of terms put forth to describe varying levels of animal or human components in media. While most of these terms began as a way to market new medium products and distinguish them from those containing serum, the result has been that without standardized definitions, it is difficult to know what components to expect in your media formulation…

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