How to Boost Your Stem Cell Culture for Cell Therapy with Cell Culture Additives

Date Published: April 11, 2025

Cell therapy is one of the most promising emerging routes to personalized medicine for the treatment of hematological malignancies and solid tumors. While cell therapy is a rather broad term with a variety of interpretations, the simplest definition is a therapy that involves the introduction of live cells into a patient’s body to treat or prevent disease. Cell therapies utilize the potential of different types of cells (including stem cells, immune cells, and somatic cells) to repair, replace, or regenerate damaged tissues or organs. Cell-based therapies may use cell transplants from donor sources, but an ever-growing arsenal of emerging therapies relies on the patient’s own cells, whether reprogrammed, genetically engineered, or otherwise manipulated.

Current Uses of Cell Therapy 1

Regenerative Medicine

Stem cell therapy is a prominent area of cell therapy. Stem cells have the ability to differentiate into various cell types, allowing them to repair damaged tissues. They are applied to conditions like heart disease, osteoarthritis, spinal cord injuries, and Parkinson’s disease.

Bone marrow transplants are widely used for treating hematological disorders like leukemia, lymphoma, and other blood cancers. Bone marrow transplants involve infusing healthy hematopoietic (blood-forming) stem cells from a compatible donor or autologous transplant to regenerate the patient’s blood and immune systems.

Cancer Immunotherapy

CAR-T cell therapy (Chimeric Antigen Receptor T-cell therapy) is a revolutionary treatment for certain types of cancers, like leukemia and lymphoma. It involves modifying a patient’s T cell to better recognize and attack cancer cells.

T-cell and NK cell therapies boost the body’s natural immune response to fight cancer by using genetically engineered immune cells.

Autoimmune Diseases

Regulatory T-cell therapy is being explored as a potential treatment for autoimmune diseases like multiple sclerosis or type 1 diabetes. The goal is to use engineered T cells to suppress the immune system’s abnormal response that attacks healthy tissues.

Gene Therapy

Gene therapy relies on genetically modified cells to correct defects or provide therapeutic benefits. For example, gene-edited stem cells are used to treat certain inherited disorders like sickle cell anemia.

Tissue Repair and Organ Regeneration

Cell therapies are being tested in conditions like heart failure and chronic lung diseases, where stem cells or other cells are injected to promote tissue repair and potentially regenerate damaged organs.

While cell therapies are relatively heterogenous in a variety of aspects -, including the employed cell type, the cell source, and how the cells are manipulated, the vast majorities of cell therapies have the common characteristic that they require the maintenance and often amplification of the utilized cells in cell culture systems in vitro.

Challenges of Cell Culture Optimization for Therapeutic Applications

Besides identifying the optimal cell type and source as well as the manipulation strategy, one of the major hurdles in the development of any given cell therapy is typically the establishment of the optimal cell culture conditions – and that frequently not once, but for multiple steps in the ex vivo cell manipulation process.

Mammalian cells have evolved over millions of years to exist within an intricate network of interwoven cells, entangled in multitudinous extracellular signals often vital for their survival, maintenance, and biological status. Cell culture of therapeutic cells not only needs to replicate that environment to maintain healthy cells, but to finely manipulate it to achieve the desired cellular changes and transitions. Using the right cell culture additives in the right amounts at the right time is paramount in the development of cell cultures for cell therapy applications.

Common Types of Cell Culture Additives Used in Stem Cell Culture

Here we want to provide an overview of commonly used cell culture additives such as small molecules, cytokines, and growth factors to optimize cell culture conditions in therapeutic applications.

Growth Factors and Cytokines in Cell Culture for Cell Therapy

Growth Factors and cytokines are diffusible signaling molecules that regulate cell proliferation, differentiation, and migration. These signaling molecules are typically secreted proteins and can act locally through autocrine and paracrine signaling, as well as endocrine through the circulatory system. Traditionally the term cytokine has been used to describe signaling molecules influencing hematopoietic cells and modulate the immune response, and growth factors was used to describe signaling molecules prominently involved in cellular differentiation, especially during embryonic development. It is becoming increasingly clear that there is pronounced overlap between these domains and the terms growth factor and cytokine can basically be used interchangeably.

Cytokines and growth factors generally act through membrane-bound receptors that relay the extracellular signals through numerous signaling pathways, including GSK-3, RAS/MAPK, PI3-Kinase/AKT and PLCγ signaling.

Cytokines and growth factors are often crucial cell culture additives for the survival and maintenance of any given cultured cell type, but their importance comes into its own when driving controlled cell differentiation in cell culture is the goal. Finding the right cocktail of cytokines and growth factors is absolute key when establishing cell therapies involving ex vivo differentiation. 2

Commonly used general growth factors:

Hematopoietic Growth Factors and Cytokines:

Small Molecules in Cell Culture for Cell Therapy

Small molecules are low-molecular-weight bioactive compounds that modulate cultured cells, by promoting cell growth, differentiation, and survival, and even reprogramming cell fate. Some combination of small molecules is typically used to modify or enhance the function of cells during the culture process to obtain cells for therapeutic applications. Small bioactive compounds play a crucial role in directing the differentiation of stem cells into the desired cell types, enhancing cell survival, and improving therapeutic efficacy. Manifold proven small molecules to enhance cell cultures exist and the untapped potential of additional compounds to boost cell therapy applications is probably even higher. To provide an overview 3 over small molecule cell culture additives, it is helpful to classify them by function:

Inhibitors of Signaling Pathways

Epigenetic Modifiers

Another category of small compounds used to modulate and optimize cell cultures for therapy applications can be classified as modifiers of epigenetic processes. Cell fate change, whether occurring in dedifferentiation into more stem-cell like state, or differentiation and maturation into more fate-committed precursors and differentiated cells, is often hindered by the inertness of the epigenetic status of the respective cell. Compounds that influence the activity of the epigenetic machinery can drastically improve the effectiveness of attempts to modulate cell status and cell fate typically required in cell therapy. The most commonly targeted cellular epigenetic actors are histone deacetylases (HDACs) and histone methyl transferases (HMTs). The following exemplary small molecule epigenetic modulators are frequently used as cell culture additives:

Metabolic Modulators and other Bioactive Compounds:

Lastly, a vast variety of small, bioactive compounds are used to enhance cell metabolism, wellbeing and survival in general through improving metabolic support and other aspects of cellular maintenance. This in theory includes basic cell culture additives for trophic support, but we will focus on less broadly used, specific enhancers of cell viability and survival with special applications:

The Right Cocktail – Of the Right Quality

Finding the right cocktail of cell culture media, media components, supplements, additives, and bioactive molecules is one of the major hurdles that needs to be overcome when developing cell culture conditions for cell therapy applications. And often, these conditions will need to be fine-tuned and adjusted throughout various steps in the de-differentiation, culturing, expansion, and differentiation of the cell type in question. In order to not only utilize the right compound, but also obtain consistent culture conditions and results, it is paramount to choose the right product quality from the start. And that is why Good Manufacturing Practice (GMP)-grade materials from an ISO certified companyare basically essential.

Summary

Why GMP Matters for Cell Culture Additives

Enzo Life Sciences is ISO 9001:2015 and 13485:2016 certified which allows us to adhere to strict GMP principles, and in turn, produce highly reproducible, high quality, and strictly controlled products that our customers can confidently insert into their highly regulated research and manufacturing efforts.

GMP-grade cell culture additives are essential for cell therapies for a variety of reasons. GMP standards help to ensure our customer’s compliance with strict regulatory standards set by bodies like the FDA and EMA. GMP guarantees traceability of materials with high quality finished product expectations by following the quality of the product from initial receipt of raw material, through the entire production process, through rigorous QC testing and release of material, and through tightly controlled maintenance of our inventoried products. GMP standards  help us to guarantee highly effective products from start to finish by  utilizing strict control over critical raw material received via a GMP compliant Receiving and Inspection processes ensuring essential specifications are met for our raw materials. Using our tightly controlled raw materials for production and by enforcing controlled production procedures, our GMP-grade additives can offer an optimal cell culturing experience which can lead to healthy and productive cells which then can ultimately provide high purity and high quality finished products for our customers.  Our GMP-grade cell culture additives can support product efficacy by ensuring that biological activity is preserved throughout our customer’s manufacturing process. Lastly, our GMP-grade reagents allow for quality maintenance during the transition of small to large scale up production efforts, ensuring that large-scale manufacturing adheres to the same rigorous standards as the smaller scale production. This guarantees smooth scaling for our customers product efforts as well as maintaining consistency across batches and safeguarding the integrity of the final product.  GMP manufacturing at Enzo Life Sciences ensures consistency as your trusted supplier, which then provides our customers with the ability to ensure that they not only have uniform product results, but experience a high degree of reproducibility.  A reproducible product is the only way to guarantee that our customers experience a smooth transition through each and every phase of their project. In summary, Enzo Life Sciences GMP-grade, high quality cell additive reagents can give researchers the peace of mind that our products can enhance and support their regulated programs from start to finish.

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Our dedicated project managers, technical
scientists, quality specialists, and application scientists deliver customized solutions you can count on.

  • Raw material qualification, testing, and tracing
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With the rise in therapeutic development, there
is a greater demand for GMP small molecules. As you advance from drug discovery to process development to manufacturing, we can help with your transition from RUO to GMP in 3-6 months.
At Enzo, we are committed to GMP. Our products are manufactured in our U.S. facilities that operates under GMP and validated in accordance with ISO 9001 and ISO 13485. Our GMP products undergo quality control testing, purity testing, and stability testing to ensure we provide the highest quality materials. We provide our customers with quality documentation and full lot traceability. Our goal is to provide unique solutions tailored to your needs and specifications. We deliver consistent and reliable GMP products to advance your manufacturing process faster and more cost-efficiently.

Hartmut Pohl, PhD

Senior Application Scientist – Linkedin profile

Hartmut is an experienced Cell Biologist with broad knowledge in neuroscience and a rare set of skills and experiences. His research career at renowned universities including the ETH Zurich and Cambridge University was focused on understanding the cell biological processes and consequences of demyelinating diseases, and was coined by multi-application approaches on the forefront of laboratory technologies. His experience with a plethora of cell biological and imaging tools is the foundation of his activities as Senior Application Scientist with Enzo, where he provides technical support and custom solutions to our customers.

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