The landscape of scientific research is undergoing a seismic shift. Researchers can no longer be content with off-the-shelf tools and materials for conducting breakthrough research. There is an increasing demand for precision, specificity, and customization. This paradigm shift is particularly evident in the realm of cell culture, where the need for tailored cell culture solutions has become as indispensable as a scalpel is to a surgeon.

At the heart of this growing demand lies the recognition that one size does not fit all when it comes to cells. These microscopic building blocks of life are, in fact, as diverse as snowflakes. Each specific type of cell is unique and has its own quirks and requirements. We cannot use DMEM, akin to a one-size-fits-all shoe, to culture cell lines like renal fibroblast. At times like these, we require Custom Cell Culture Media that will provide sustenance to these cells with optimized nutrient levels. They offer a tailored environment that caters to the specific needs of each cell type.

In this blog, we will advocate to you how custom cell solutions can help expedite your research, leading to biologically significant results in humans.

Why do we need custom cell culture media and custom cell solutions?

There are various reports that show that media components can significantly affect cell growth, expression profile, transfection efficiency, surface marker expression, and physiology of the cells.

A study published in “THE CELL” in 2016, demonstrated how L-arginine concentrations can impact the metabolic profile and survival of T cells. Another study showed that adding cytokine IL-21 in the cell culture media can significantly improve the transfection efficiency of T-cells.

These studies are just a glimpse of what the literature is telling us about the importance of custom cell culture media. Another level of complication that is often overlooked is the presence or use of primary cells that are required for studies. For instance, you are studying T-cells biology but if you are using immortalized cells, how will they be able to give you results that are similar to physiological conditions? Thus, there is a need for custom cell solution services, which can provide you with human primary cells from healthy donors or donors with specific conditions.

It is a time-consuming process to find a donor, collect the sample, and then expand the primary cells for your experiments. This is all needed to be done while you have to do regular research work. It just creates unnecessary stress for you and your research team. So, what is the solution? It is important to put your trust in the primary cell manufacturers.

Kosheeka: Your Partner in Cell Culture Customization

We at Kosheeka have over a decade of experience providing Custom Cell Solutions to researchers and contract research organizations. Our custom cell culture media are not merely mixtures of chemicals in water; they are meticulously crafted formulations for your cells. Our media are made with special attention to the cells’ specific nutrients, pH levels, osmolarity, sterility, and endotoxin levels.

On the other hand, our custom cell culture media is akin to providing the cell with a carefully balanced, nutrient-rich soil, ensuring optimal growth and vitality. We follow a comprehensive customization plan that is flexible. We work with you to understand your specific requirements, overcoming any challenges that may arise. Our team of scientists is committed to transforming your research aspirations into reality.

With our deep-rooted passion for cellular biology, we are well-equipped to provide you with rare primary cells, develop novel cell lines, or help you create intricate experiments.

Benefits of using custom cell culture media and custom cell solutions

The advantages of using custom cell culture media are manifold. These include:

• Scale-up of the custom recipes with top-quality powdered media.

• Media components are isolated from qualified sources, ensuring GMP-compliance.

• Quick availability of primary cells for your experiments.

• Flexibility to customize existing media for your cell culture experiment.

• Comprehensive regulatory documentation for the cells used in treatment.

• Complete quality check reports to ensure that the cells and media you receive boost cell growth.

Conclusion

Custom cell solutions are quickly becoming the need of the hour as we dive deep into understanding human biology. Using primary cells enables us to make groundbreaking discoveries and develop drugs for therapeutic purposes. Custom cell culture media is crucial for providing the right environment that accelerates cell growth while maintaining physiological similarities with human body cells.

With a decade of experience in providing high-quality custom cell solutions, Kosheeka is in the perfect position to ensure that your research is ready to go to the next level. 

For more information, please contact us at +91-9654321400 or visit our website. www.kosheeka.com

Cardiovascular disorders have been the predominant cause of mortality on a global level. It has urged the scientists to develop new treatment procedures and therapeutics. However, it requires understanding of cardiac cellular pathways and its genotype. Moreover, the evaluation of the safety and efficacy of the drugs necessitates an appropriate in vitro and in vivo model. Pigs are almost 98% identical to humans and are already employed for in vivo research. Therefore, the employment of primary swine or porcine cardiomyocytes from pig for in vitro studies has clinical relevance in cardiac research.

Why Primary Swine Cardiomyocytes?

Cardiomyocytes are the muscle cells present in the heart and are responsible for its contraction. Human cardiac muscle cells are the most appropriate in vitro model for cardiovascular research. However, isolation of cardiac tissue from humans has limited availability. Cardiomyocytes developed from induced pluripotent stem cells have been employed. But they are immature and differ from adult human cells in structural and functional aspects. Another feasible option is Swine Cardiomyocytes.

Human and pig hearts bear resemblance in hemodynamic, structural, electrophysiological, and size-based characteristics, prompting the use of porcine cardiomyocytes in preclinical research. Cardiac features such as heart rate and contractility are extensively studied in research and are comparable between humans and pigs. The genetic profiles of human and pig cardiac muscle cells are also similar. These parallels between human and pig hearts have prompted the use of primary porcine cardiomyocytes as an in vitro model for cardiovascular research.

Culture of Primary Swine Cardiomyocytes

The primary porcine cardiomyocytes have generated interest in outlining their isolation and culture process. The perfusion method with Langendorff apparatus and the following enzymatic digestion is used for isolation of the cells. The culture of primary cells aims to maintain a homogenous cell population for the long-term. Their culture media comprise a basal medium of either DMEM and F-12 or M-199. Initially, cells show negligible attachment to the substrate in the serum-based medium. Lack of attachment caused them to assume a rounded morphology. However, they transformed to their original shape as they adhered to the substrate. This culture method was termed the redifferentiation. Furthermore, attachment to the substrate didn’t require serum in the media. This culture method is the rapid attachment method. It served as a better method for retaining the morphology and functions of cardiac muscle cells. The medium also requires additional supplementation with epidermal growth factor (EGF).

Role of Primary Swine Cardiomyocytes in Cardiovascular Research

With the rising burden of cardiovascular disorders, the research into cardiac tissue structure and function has increased. The porcine cardiomyocytes have found applications in cardiovascular research owing to their abundance and cost-effectiveness. Researchers can investigate the underlying signaling mechanisms behind the electrophysiological and muscular activity of cardiac tissue. It has prompted the development of new therapeutic interventions. Scientists have also genetically engineered these cells to facilitate their xenotransplantation. Experimenting on them has provided insights into the development and regeneration of the heart. It has aided in the development of treatments to repair cardiac injury. The efficacy of these drugs could be evaluated on these cells for their effectiveness. Primary porcine cells have the potential to form 3D models that can imitate the in vivo environment of the organ.

Applications of Primary Swine Cardiomyocytes in Drug Development

The pharmaceutical research focuses on the formulation of new medications and improving the already existing drugs with regard to reducing their cost and off-target effects. The cardiac issues have various comorbidities. Frequently the therapeutics used as standard treatment of care, such as kinase inhibitors, antibiotics, antidepressants, etc., also exhibit cardiotoxic effects. Several drugs have been withdrawn from the market for causing cardiac issues. It has urged the researchers to perform toxicological screening of the newly developed drugs on cardiomyocytes. But toxicity screening was performed on kidney cell lines. However, they lack the various ion channels and the signaling pathways that are an integral part of the human heart. The evaluation of drugs on kidney cell lines even resulted in removal of drugs during the clinical trial stage due to false toxicity issues. Primary porcine cardiac cells are more suitable for screening drugs for triggering cardiac-related toxic effects.

Conclusion

The growing cases of cardiovascular disorders have encouraged the scientific community to devise therapeutic alternatives. Every new-age therapeutic undergoes rigorous testing in animal models. Small animal models, especially rodents differ in their heart rate, life span, surface-to-body ratio, and genetic profile from the human heart. For cardiovascular research, pigs are the suitable model since porcine heart share similarities with that of the human heart. Additionally, they are easily available and are cost-effective. These cells are also suitable for toxicity assessment of non-cardiac-related drugs. Scientists have gained significant insights from in vitro research on primary porcine cardiomyocytes. They have wide applications in basic, pharmacological, and toxicological research. Therefore, they would be beneficial for research into cardiac tissue. Kosheeka delivers high-quality cardiomyocytes from the ventricle of adult porcine heart at passage 2.

Mesenchymal stem cells (MSCs) have applications in the treatment of diverse disorders. They were first identified in bone marrow. However, their extraction from bone marrow was a complicated process, which fueled the research on discovering other MSC sources. Over the years, different sources of MSCs have been found, such as umbilical cord, adipose tissue, and dental pulp. Among them, adipose tissue has gained preference for its easy isolation and higher availability. Adipose tissue provides a much higher cellular yield than bone marrow, which is beneficial for research and therapy purposes. Therefore, numerous clinical trials have utilized adipose-derived MSCs for treating neurological disorders, cardiovascular issues, bone regeneration, diabetes, etc. 

Human Adipose-derived Mesenchymal Stem Cells (AD-MSCs)

Human Adipose-derived Mesenchymal Stem Cells are multipotent adult stem cells present in the adipose tissue located in the visceral and subcutaneous regions. The term to define this cell population varies from AD-MSCs to adipose stem cells (ASCs) or adipose-derived stromal cells (ADSCs). In comparison to bone marrow MSCs, adipose tissue cells demonstrate longer lifespan in culture and higher proliferative capacity. Their differentiation expands further from MSC-lineages to hepatocytes, epithelial cells, neurons, cardiomyocytes, pancreatic islets, and endothelial cells. 

The extraction procedure involves liposuction surgery or lipoaspiration- a minimally invasive, safe, and cost-effective procedure. The amount of stem cells in the adipose tissue aspirate is also higher than that of the bone marrow aspirate. Studies have calculated that per mL marrow aspirate comprises 0.001-0.01% stem cells, whereas the percentage in adipose tissue is 1-10%. It gives an approximate value of 1 × 10^5 stem cells per gram of tissue. 

Immunomodulation by Adipose Stem Cells

Self-renewal and multi-lineage differentiation have been the defining features of AD-MSCs. In addition, these cells also possess immunomodulatory properties, that is, they secrete cytokines to regulate immune response. According to a few studies, they release an increased number of cytokines in comparison to their bone marrow counterparts. Additionally, they release prostaglandin E2 (PGE2) and IDO, which have immunomodulatory properties. For instance, PGE2 induces macrophages to produce IL10 to inhibit NK cells and T helper lymphocytes.

Furthermore, AD-MSCs lack CD80, CD86, MHC II, CD40, CD40L, etc. which suppresses T lymphocyte activation during allogeneic transplant. They also inhibit the proliferation and differentiation of B lymphocytes, which prevents graft-vs-host disease. While the absence of specific surface proteins on AD-MSCs suppresses immune response, the presence of proteins on HLA-G5, PDL1, and galectins induces immunogenic tolerance. HLA-G5 inhibits dendritic cells, natural killer (NK) cells, and T lymphocytes. PDL1 and galectin are immune checkpoint regulators that subdue immune responses.

Factors Affecting Immunomodulation

The immunosuppression ability of AD-MSCs is remarkable. These cells can induce the proliferation of CD5+ B regulatory lymphocytes. However, this ability is dependent on several factors. Studies have reported that at late passage, the expression of MHC II, CD80, and CD86 upregulates in AD-MSCs, driving T lymphocyte proliferation. Another research inferred that AD-MSCs from female donors are more potent immunomodulators than those of male donors. It even showed the gender-based difference in the molecules that regulate immune response. 

Moreover, research has found that dosage and timing of infusion also alter their properties. Many studies have also demonstrated that the cellular process and yield are influenced by the location of the tissue used for cell extraction. For instance, there is reduced apoptosis in abdominal AD-MSCs. These findings imply that donor features, treatment regimen, and extraction site affect the consistency of the therapeutic effect.

Enhancing Immunomodulation

Different approaches are under investigation for improving the immunomodulatory property of AD-MSCs. A study demonstrated that IFNγ produced by activated T lymphocytes primes AD-MSCs against T lymphocyte proliferation. Therefore, preconditioning with diverse inflammatory mediators such as IFNγ, TNFα, IL1, IL2, etc. can augment the ability to govern immune response. For instance, IL1β-primed AD-MSCs can suppress joint inflammation. A different concept is to retain the immunomodulation ability of AD-MSCs through their 3D culture. The spheroid culture has exhibited increased efficacy in controlling immunity. Moreover, their genetic manipulation can also confer heightened immune regulation. Integrating hypoxia-inducible or inflammation-sensitive promoters can trigger cytokine secretion in specific disease signals. 

Conclusion

Adipose tissue has been an attractive source of MSCs. The minimally invasive extraction procedure and high cell yield have fueled its applications in clinical trials. Their immunomodulatory properties have therapeutic potential for autoimmune and inflammatory disorders. Additionally, this property also prevents immune rejection and GVHD after allogenic transplant. Moreover, this property can be augmented through different strategies for better efficacy.

However, it should be noted that their therapeutic ability might vary with different factors, including donor attributes. It could contribute to the different responses often observed in clinical trials or research studies. AD-MSCs belonging to different donors are also affected by age and environment. Therefore, considering these factors in mind before their isolation can maximize the consistency of the cells. Advancells maintains the consistency standards while offering Human Adipose-Derived Mesenchymal Stem Cells at early passage with relevant documentation of the donor.

In vitro culture has contributed substantially to biomedical research. However, the availability of primary and cancer cells has complicated the choice between the two cell types. The key hurdle in primary cell culture is their establishment in the in vitro environment. The challenging process has led to the emergence of research companies' primary cell developing services. These are private sector investments that contribute to scientific growth. They enable innovations by providing primary cells and saving researchers the hassle of the isolation process.

Primary Cells versus Cell Lines

In 1907, Ross G. Harrison successfully cultivated frog embryo nerve fibers in the in vitro environment, marking the beginning of tissue culture. Another milestone was achieved when HeLa cells, belonging to cervical cancer patient Henrietta Lacks, showed unlimited growth in the tissue culture laboratories. It inspired the development of cell lines with indefinite proliferation within laboratories. The available choices sparked the debate on primary cells vs cell lines. Before selection, the following distinctive properties of both choices should be kept in mind.

Origin: Primary and cancer cell population belong to non-malignant and malignant tissues, respectively. Cell lines also develop from immortalization of Primary Cells by gene transfection or viral infection, which dysregulates the cell cycle. Additionally, telomerase (hTERT) transfection prevents telomere shortening and apoptosis conferring immortalization.

Lifespan: The proliferation ability is the key deciding factor in the selection. Primary cells live up to a few passages and undergo apoptosis. On the contrary, cancer cells proliferate for an indefinite period. Both of them encounter genetic drift and deviate highly from the original population with each subculture.

Tissue Characteristics: The basic difference between the two choices is their tissue characteristics. Primary cells resemble the in vivo tissue characteristics, but with every subculture, they incorporate changes, reducing the similarities with the tissue. Cancer cells retain the properties of malignant tissue, whereas cell lines have the least resemblance to the tissue.  

Homogeneity: Research emphasizes on reproducibility which in turn requires consistency in cells. However, population and tissue demonstrate variation that is reflected in cells. Primary cell population is pooled from several donors and thus constitute a heterogeneous population. Cancer cells show the tumor tissue heterogeneity but in vitro cultures immortalized in the laboratory are relatively homogenous.

Advantages vs Disadvantages

The difference between the cell types grants them advantages and disadvantages that can aid in the choice between them.

Culture: The finite life span of primary cells makes their in vitro culture difficult. They require growth factors and a medium specific for their survival and proliferation. The short lifespan also limits the cell quantity available for assays, mandating repeated procurement or isolation. Cancer cells are typically easy to cultivate in vitro with minimal requirements for optimal growth. They are available in sufficient quantities to conduct multiple assays.

Relevance: Similarities with the tissue provide more accurate results which are reproducible in the in vivo models. Thus, primary cells have established their significance over the other options. Cancer cells have applications in oncology research, but cannot be exploited for other fields.

Weighing the advantages and drawbacks can assist in making a suitable selection. The lack of donors and the need for surgical procedures for isolation, together with limited life span, make primary cell culture difficult, redirecting the focus on other alternatives. Few of them only live up to five or fewer passages, either undergoing differentiation or apoptosis. Therefore, the alternatives gain preference as suitable in vitro models for long-term studies.

Primary Cell Developing Services

The establishment of primary cell culture is a complicated process that demands optimization at every step. It entails investment in terms of money and time. A research laboratory has limited resources to spend in this direction without the assurance of quality. Their budgets and deadlines often redirect them towards Primary Cell Developing Services. These are private sector ventures focused on manufacturing these cultures with high quality. 

Kosheeka is one such venture that incorporates an expert team of scientists and streamlined workflows. They result in short turnaround times. Its regulatory compliance and advanced facilities demonstrate its dedication to culture services. It offers custom solutions that align with the research needs of the scientists. Its experts also guide the researchers for the in vitro culture process and any issues faced during experimentation.

Final Thoughts

The selection between the cell types for research have been subjected to debate for years. The choice boils down to cost constraints and study design. Low budget promotes the use of immortal populations, whereas future applications in the in vivo models underscore the use of primary cells. The extraction and culture of the latter is a complex process beyond the limits of the research laboratory. Research companies facilitate scientific explorations by establishing their cultures and offering them at reasonable charges. It reduces the financial burden on researchers, saving valuable time and efforts. Kosheeka is a cell manufacturer company with a diversified inventory of cells and its products, delivered with assured quality to elevate biomedical research.