In Vivo vs. In Vitro Differentiation: In Vitro T Cell Differentiation Protocol and Assays

In vitro cell differentiation and cell separation assays are easier to perform and offer greater control in experimentation compared to their in vivo counterparts, making in vitro assays especially useful in the early, pre-clinical phases of therapeutics research. Leveraging innovative microbubble technology, the T cell isolation products developed at Akadeum Life Sciences provide an affordable, accessible, and accurate method for conducting in vitro cell separation, making cell differentiation protocols more efficient than ever before.

In vivo vs in vitro Assays

Biological sciences and pharmaceutical research involve both in vitro and in vivo assays to study specific biological mechanisms and analyze the efficacy of experimental treatments. The term in vitro (Lat. “in glass”) refers to procedures that occur outside of a living organism, typically in test tubes, petri dishes, or other isolated laboratory environments. Meanwhile, in vivo (“within the living”) procedures take place within a living organism, organ, or cell culture.

Each process has unique advantages and drawbacks that make it suited for specific phases of research. In vitro assays that don’t require living test subjects are cheaper to run, require less specialized training, and allow scientists to isolate the mechanisms-in-question from the complexities of a living environment. Meanwhile, in vivo assays within a living organism grant a fuller picture of the biological systems in-question and what effects a given treatment will have on the organism.

In vitro Cell Differentiation

In general, cellular differentiation describes the process in which a cell changes from one type to another, and commonly refers to the transformation of stem cells into more specialized cells. Studying stem cell differentiation allows researchers insight into how specific cell-types or complex biological structures develop. Research involving in vivo and in vitro differentiation assays show that while, in vitro cell differentiation protocols alone are not sufficient to predict how differentiated cells will behave within the complex environments of a living organism. Still, in vitro assays play a crucial role in understanding the fundamental mechanics of cell differentiation, as they provide an accessible and cost-efficient method for testing and optimizing therapeutics treatments before risking the health of living subjects in in vivo trials.

Immunologists rely on in vitro T cell differentiation methods to study the process in which Hematopoietic Stem Cells [HSCs] migrate from bone marrow to the thymus gland, where they then develop into a variety of distinct T cell types to support the immune system:

  • TMEM –memory T cells recall effective immune system responses when they re-encounter pathogens or familiar infections
  • TREG –regulatory T cells distinguish between foreign or infected cells and the body’s own healthy cells
  • TH –helper T cells secrete proteins called cytokines that signal B cells to differentiate into blood plasma cells that produce antibodies
  • TC –cytotoxic or killer T cells bind to proteins on infected cells and release cytotoxins that induce cell death in their targets

After maturing in the thymus, T cells circulate through the bloodstream and lymphatic system to hunt pathogens or keep watch over specific regions of peripheral tissues throughout the body. Because in vitro studies occur outside living environments, researchers are able to exercise greater control in manipulating the T cell differentiation process, making in vitro T cell differentiation protocols essential both to advancing scientific knowledge and developing therapeutic treatment of immunological disease.

Cell differentiation also occurs in Embryonic Stem Cells [ESCs], which can develop into any type of cell (aside from placental cells), and studying these processes is critical to the development of clinical therapies and treatments for a wide range of conditions from genetic disorders to diabetes and heart disease. While ethical concerns about the health of living embryos as well as technical and resource limitations constrain the efficacy of in vivo ESC differentiation assays, in vitro differentiation of Embryonic Stem Cells is affordable, requires less specialized training, and allows researchers an increased ability to manipulate the cell differentiation process with no risk to living test subjects.

In vitro Cell Separation

In order to perform cell differentiation assays, scientists must first isolate the cells in question. Cell separation (also called “cell isolation”) describes processes for targeting specific cell types within a biological sample and setting them apart from the mixture for targeted analysis and experimentation. By isolating specific cells from a heterogeneous sample, scientists can study their characteristics to learn about their behaviors within the human body and develop treatment applications in fields including immunology, oncology, and stem cell therapy.

Such cell separation is a featured step in many cell differentiation assays, wherein cells whose differentiation are to be studied are targeted and isolated in order to probe the mechanisms on which they can be coerced to differentiate. Given the difficulty of isolating specific cells within a complex biological system in vivo, researchers often favor in vitro cell separation methods in order to optimize control, cost- and training-efficiency, and both purity and viability of cellular health for downstream study and treatment applications.

T cell isolation is essential to immunological research and involves separating T cells from blood or tissue samples for analysis and development of immune system treatments. Once separated from their sample, T cells can then be genetically modified to target and attack specific pathogens, viral infections, and even cancer cells. T cells are typically isolated outside of living subjects, as in vitro T cell activation grants researchers precise control over the expression of regulatory and signaling proteins such as T cell activation cytokines that tell other immune system supporting B cells to differentiate into antibody-producing blood plasma cells. Furthermore, working in vitro allows researchers to make genetic modifications to a patient’s T cells after they have been isolated from a blood sample, tweak the T cells to combat specific pathogens, and then re-introduce them to the patient—such in vitro T cell separation and modification protocols are essential to immune system treatments such as CAR T cell therapy.

In vitro Cell Isolation for Cell Differentiation Assays

Although in vivo assays present a more-complete look at the complexities within an organism, these methods are expensive and difficult to perform with precision. Meanwhile, in vitro procedures are more accessible, easier to employ, and offer greater control over the cell isolation or cell differentiation processes. In fact, in vitro protocols are particularly effective for conducting T cell differentiation or T cell separation assays, as well as patient-specific T cell therapeutics treatments.

Akadeum Life Sciences offers a wide range of affordable and easy-to-use in vitro cell isolation kits that utilize innovative microbubble technology to isolate a variety of cell types. Akadeum’s Buoyancy Activated Cell Separation [BACS] process gently and efficiently separates even the most fragile target cells from a sample while ensuring high purity, yield, and viability of enriched cell populations at-scale. Check out Akadeum’s line of BACS cell isolation products to optimize your own cell differentiation assays!

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