#cell lines

[secondwheel]

An immortal cell line

[sciencenewsforstudents]

How making a COVID-19 vaccine confronts thorny ethical issues

COVID-19 vaccines will face plenty of ethical questions. Concerns arise long before anything is loaded into a syringe.

Ethical concerns abound in the race to develop a COVID-19 vaccine. How do we ethically test it in people? Can people be forced to get the vaccine if they don’t want it? Who should get it first?

Tackling those questions demands that a vaccine exist. But a slew of other ethical questions arise long before anything is loaded into a syringe. In particular, some Catholic leaders in the United States and Canada are concerned about COVID-19 vaccine candidates made using cells derived from human fetuses aborted electively in the 1970s and 1980s. The group wrote a letter to the commissioner of the U.S. Food and Drug Administration in April, expressing concern that several vaccines involving these cell lines were selected for Operation Warp Speed — a multibillion-dollar U.S. government partnership aimed at delivering a COVID-19 vaccine by January 2021.

The group urged the FDA to instead provide incentives for COVID-19 vaccines that do not use fetal cell lines. But, as virologist Angela Rasmussen of Columbia University pointed out on Twitter, those other vaccines are being developed with scientific input from research using HeLA cells — which come with their own thorny ethical issues of consent.

Here’s how scientists and bioethicists are thinking about the cell lines they use as they develop COVID-19 vaccines.

[bpod-bpod]

Signature of Sensitivity

When it comes to treating cancer, one size doesn’t fit all. Each patient’s disease is a genetic pick-and-mix of alterations and mutations that affect how it behaves and whether it's resistant or sensitive to various types of chemotherapy. To find out more about how genetic variations within tumours influence treatment, researchers have been testing common chemotherapy drugs on more than 200 genetically distinct cancer cell lines growing in the lab. Each column in the grid represents one cell line, tested with 21 different drugs – one for each row. Blue and green highlight sensitive cells that are easily killed, while red and pink patches carry genetic alterations making them resistant to that particular treatment. By combining all this data together, scientists can generate a set of ‘signatures’ that can be used to figure out how different drugs work and identify treatments that are likely to be effective for individual patients.

Written by Kat Arney

Trick Or Treat: Probing Cancer's Response to Chemotherapy; A winner in the 2020 Koch Institute Image Awards

Image by Peter Bruno, Aslı Gökdemir, Ryan Hayman and Michael Hemann; Hemann Lab MIT

Image courtesy of the Koch Institute Image Awards

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[drawittoknowitmedical]

CMP:

Megakaryocytes:

In the presence of interleukin 11, the descendants of the CMP gives rise to megakaryoblasts, which are large basophilic cells with a bean-shaped nucleus.

Under the influence of thrombopoietin, the megakaryoblast matures to form the megakaryocyte (be aware that thrombopoietin is also referred to as THPO and megakaryocyte growth and development factor (MGDF), and c-Mpl ligand).

Megakaryoctes are large cells with multilobed, irregular nuclei.

The plasma membrane of mature cells invaginates to form demarcation membranes that fragment the cytoplasm.

The fragments are shed into the blood as platelets, aka, thrombocytes, which perform key hemostatic functions.

Erythrocyte:

Erythropoietin (EPO) initiates the erythroid series, which begins with formation of the proerythroblast and ends with the mature erythrocyte (aka, red blood cell).

Granulocytes:

Granulocyte-macrophage colony stimulating factor (GM-CSF) triggers formation of the myeloblast, which will give rise to 4 of the 5 white blood cell lines:

Eosinophils

Interleukin 5 triggers production of eosinophils

These white blood cells are characterized by multi-lobed nuclei and bright orange/red staining granules.

Basophils

Interleukin 3 triggers production of basophils, which, as their name implies, are highly basophilic. (be aware that interleukin 3 has widespread influence over the other cell lines, too.)

Neutrophils

Presence of granulocyte – colony stimulating factor (G-CSF), myeloblasts become neutrophils.

These white blood cells comprise lobulated nuclei and a pale pink cytoplasm.

Agranulocytes:

Monocyte

In the presence of monoblast – colony stimulating factor (M-CSF), the monoblast gives rise to the monocyte

This white blood cell has a bean-shaped nucleus, with visible chromatin, and blue-gray cytoplasm.

Outside of the red marrow, the monocyte differentiates:

In the connective tissues, it becomes a macrophage, which can phagocytose dozens of particles, even damaged red blood cells.

In the bone, the macrophage becomes an osteoclast, which resorbs bone tissue during growth and remodeling.

[biomedicalscientist]

Development of the female breast (cell lines)

[kosheeka]

How To Select Cancer Cell Lines For Your Experiments?

Before choosing cancer cell lines for your experiments, it’s essential to evaluate a few critical factors. Selecting the right cell line impacts the reliability and relevance of your results. Here are five key questions to consider to ensure that your chosen Cancer Cell Line is credible, contamination-free, biologically appropriate, genetically stable, and suitable for your specific experimental requirements.

Here is a list of a few questions that you must ask before selecting cancer cell lines for your experiments:

1. Is it a credible cell line? 

It is of utmost importance that you ask the question of whether a certain line is what it is claimed. You would be shocked to know that there are over 500 cell lines that have been masked and misidentified in the Register of Misidentified Cell Lines (v13 April, 2024). You sure can obtain cell lines from third-party cell banks like Kosheeka and NCCS. Even when you obtain these cell lines it is best to have them annually assessed using STR profiling.

If you start an experiment with a commercial cell line like BCC1/KMC, it could be you are actually not working with them. The only real way for you to make sure that you actually pick up cell lines that you need is to always go for a certificate of authentication and do STR profiling. STR profiling might be a long process, but doing it is necessary to ensure that you have an authentic biological model that works as it is intended.

2. Is it free of contamination?

Yes, even if you get an authentic line, it is necessary to ensure you have contaminant free cell lines. A simple bacterial infection can put added stress on these cell lines that will make their biology change. Now, if you get an infection that alters the biology, your experiments will change. That's because the microorganisms are so small and don't look like other bacteria (they don't have a solid wall, instead they have a plasma-like shape).

Reliable cell providers will provide cell stocks confirmed to be mycoplasma-free. When the contamination status of a cell stock is uncertain, it is advisable to cultivate it in a quarantine setting and confirm its "clean" status before transferring it to public circulation. This approach minimizes the chance of mycoplasma contaminating a whole cell culture laboratory.

3. What is the passage number of cells?

For in vitro cultures you will notice that their is genetic instability, and Cancer Cell Lines start to accumulate these genetical errors. Even a recent study from Domcke et al., found "pronounced differences in molecular profiles between commonly used ovarian cancer cell lines.This is likely attributable to the age and resultant prolonged culture of particular lines, resulting in their divergence from the original malignancies.

To mitigate this, it is advisable to sustain cell stocks at the lowest passage feasible and to routinely (every 2-3 months) start cell culture with a new vial. Temporal alterations in cell lines may, in some instances, be visibly discernible; for instance, prolonged culture of U87 cells might result in a diminished monolayer and the formation of spheroids from the flask surface.

4. Does it show the right biology?

Your choice of cell line will almost definitely be determined by the question or problem you're attempting to address. If you're studying a certain disease condition, the closer the cell line resembles that sickness, the better.

Even across illness kinds, careful selection is essential. For example, if you're studying breast cancer, you might be interested in only triple negative breast cancer, or you might want to look at a variety of subtypes. Your research objectives will guide your choices.

If you are studying a disease, the Cancer Cell Line Encyclopaedia (CCLE) is a fantastic resource for identifying a cell line that expresses a specific biomarker or has a specific genetic abnormality. The CCLE offers public access to genomic data, analysis, and visualization for about 1000 cell lines, allowing you to not only choose an appropriate cell line but also place any results you generate within the context of the CCLE's enormous data set. 

If your focus is on basic biology rather than disease biology, you will frequently come across a model line from which a body of work has been created. By taking this same approach, discoveries can be placed in the context of existing scientific knowledge.

This method of selection, however, produces a self-perpetuating bias toward published cell lines, which may not have been chosen because they are the best suitable. Unsurprisingly, the age of a cell line can influence how frequently it appears in the literature, as with the first immortalized cell line, HeLa cells, which have been widely employed (their simplicity of cultivation no doubt contributing to their popularity).

The best method is to identify numerous cell lines and show results over a panel. This not only increases confidence in the results, but it also eliminates the idea that they are simply a quirk of a specific line.

5. Is it suitable for your experiment?

The selection of a cell line for any experiment necessitates a balance between identifying the most physiologically relevant model and opting for a cell line that is manageable for experimentation.

Cultural conditions, growth rate, transfectability, morphology, subcellular structure (for microscopy), and cost significantly influence the studies conducted and their probable success.

Induced pluripotent stem cells (iPSCs) may provide a more "normal" model compared to cancer cell lines; however, the demanding cell culture protocols and related expenses may render these lines unsuitable for certain laboratories.

The objective is to identify the optimal compromise—a cell line that has comparable biological characteristics, facilitates rigorous and efficient scientific inquiry, and withstands validation in external laboratories.

Detailed culture conditions and growth rates are provided by Kosheeka for numerous widely used cell lines. 

[kosheeka]

Differences and Advantages :Immortal Cell Lines and Human Primary Cells

What are the differences and advantages of using immortalized cell lines & primary human cells? For more info, Contact us at [email protected]

When you are starting your cell culture research, the primary requirement is having cells, and deciding whether human primary cells or cell lines will be beneficial, is a major task!

Read the blog to learn about the differences and advantages of cell lines and primary cells:

[rootsanalysis-blog]

CHALLENGES AND OPPORTUNITIES FOR CELL LINE DEVELOPMENT AND CHARACTERIZATION SERVICE PROVIDERS

In the past few years, there has been a significant surge in the demand for recombinant proteins and cellular systems for biomedical research, diagnostics and different therapies. In order to support the large- scale production and manufacturing of such cell- based products, there is an imperative need for the efficient development of high- quality stable cell lines. In fact, 60-70% of manufacturing processes of available biologics require live mammalian systems. A cell line refers to a defined population of cells that originates from a single common ancestor cell and have the ability to retain stable phenotypes and functions.  In general, once the primary culture is sub-cultured, it becomes a cell line

 To request a sample copy / brochure of this report, please visit : https://www.rootsanalysis.com/reports/cell-line-development-and-characterization-services/request-sample.html

 Cell lines have revolutionized scientific R&D and are currently being used for a myriad of applications.

 The development and characterization of cell lines is both technically challenging and financially demanding. It requires cost intensive materials, specialized genetic engineering technologies, aseptic cell culturing conditions, specialized infrastructure for cell storage, and product-specific analytical techniques for cell / cell line testing. Recent years have witnessed the emergence of a large number of highly qualified contract research organizations (CROs), contract manufacturing organizations (CMOs) and contract development and manufacturing organizations (CDMOs) that assist drug developers and researchers, and offer the following benefits:

§  Reduction of economic burden: Cost of developing and characterizing cell lines may vary between few thousand dollars to hundreds of thousands of dollars, depending on the cell line engineering methods, expression hosts, upstream and downstream process development and the type of tests required during research and / or the production phases. For instance, cell lines edited using CRISPR/Cas9 system are likely to be priced higher. Service providers have the potential to reduce the financial risk for the stakeholders that do not have the capacity, capability or the in-house expertise to carry out such operations.

§  Enables companies to focus on research: Contract service providers offer a variety of services for different cell lines, allowing developers and researchers to avail analytical tests as per their requirements. Therefore, outsourcing cell line development and characterization operations allows product developers to focus their efforts on R&D activities.

§  Knowledge expertise: Service providers have the necessary skillset to carry out complicated procedures in minimum time with reduced chances of failure. In addition, these organizations often provide consulting services to assist researchers to obtain accurate results and address regulatory concerns.

§  Risk Sharing: One of the crucial factors determining the net outcome of any industrial process is risk-analysis. Outsourcing few of the components of an industrial process is just like shifting certain responsibilities to the outsourced vendor. Henceforth, it is a mutual understanding between the companies to handle the risk factors in a better way.

§  Integrated offerings: Nowadays, contract services providers are offering full range services, from cell line generation to screening and characterization, including the establishment and maintenance of master cell banks (MCBs) and working cell banks (WBCs), along with the end-product formation from cell lines.

 For additional details, please visit : https://www.rootsanalysis.com/reports/cell-line-development-and-characterization-services.html

 Despite the increasing opportunity in this domain, high entry costs (associated with building the required technical expertise and setting up of facilities) and rising regulatory stringencies, are the primary impediments to the number of stakeholders entering this market. For sponsors looking to outsource, there are multiple areas of concern associated with engaging service provider entities; some of the prominent challenges faced, include lack of necessary expertise for substandard cell growth troubleshooting, and process transfer related complexities. The selection of an inappropriate service provider partner can prove to be disastrous in the long run, creating issues, such as delays and cost overruns.  

  Despite the aforementioned challenges, the growing demand for novel biologics, especially amidst COVID-19 pandemic, impending patent expiries of several blockbuster biologics, growing popularity of regenerative medicines and cellular vaccines, and advancements in genetic engineering and bioanalytical technologies, are some of the major factors that are anticipated to drive the growth of cell line development and characterization services market, in the mid to long term.

 Key Questions Answered

Who are the leading players     offering cell line development services?

What kind of CDMO support is     available for cell line development, across different regions?

What are the common sources, gene delivery     methods, protein yield and affiliated services offered by the cell line     development service providers?

Who are the leading industry and     non-industry players offering cell line     characterization services?

What are the most popular services     offered for characterization of cell lines?

Which partnership models are     commonly adopted by stakeholders in this industry?

How is the current and future     opportunity likely to be distributed across key market segments?

What are the anticipated future     trends related to cell line development and characterization market?

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3.      HER2 Targeting Therapies Market, 2021-2030

  About Roots Analysis

Roots Analysis is one of the fastest growing market research companies, sharing fresh and independent perspectives in the bio-pharmaceutical industry. The in-depth research, analysis and insights are driven by an experienced leadership team which has gained many years of significant experience in this sector. If you’d like help with your growing business needs, get in touch at [email protected]

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