Applications

Primary Cell Culture

What are stem cells?

Stem cells are essential for all living organisms due to their unique regenerative properties. When adult stem cells divide, they have the potential to develop into many other types of cell with more specialized functions, such as muscle cells, brain cells or red blood cells. This remarkable multipotent differentiation ability means that stem cells underpin many aspects of early development and growth. In some adult tissues, such as brain, muscle and bone marrow, discrete populations of adult stem cells can also replenish other cells that are lost through normal wear and tear, injury, or disease.

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Three characteristics that distinguish stem cells from other cells

They are unspecialized cells that can differentiate into specialized cell types
They can divide and renew themselves for long periods
Under certain physiologic or experimental conditions, stem cells can be induced to become tissue- or organ-specific cells with special functions

These characteristics mean that stem cells have many exciting applications in disease research and in developing novel therapeutics. For example, in regenerative medicine, work is well underway to understand how stem cells can be used in cell-based therapies to treat a range of diseases. In vitro stem cell culture is also being used to screen new drugs as well as to develop model systems for studying pathological pathways of disease to identify therapeutic targets.

Scientists have primarily worked with embryonic or pluripotent stem cells (involved in early development) and adult stem cells (involved in repair) derived from both animals and humans. A new type of human stem cell called induced pluripotent stem cells (iPSCs) has also recently been developed. These are genetically reprogrammed, specialized human adult cells that assume a stem cell-like state. The reprogramming process, known as transfection, has been previously challenging in stem cells, particularly using non-viral transfection methods.

In the following sections, we outline the methodological principles of in vitro stem cell culture and its benefits and challenges, and how our innovative Nucleofector^® Technology can help to overcome the challenges of transfecting stem cells. We also review the different types of stem cells we offer, including iPSCs and adult stem cells such as mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs). This will help you to identify which types are best suited to your needs. Lastly, we look at the different applications of stem cell culture, including in disease research and therapy, to elucidate the importance of stem cells in clinical and research progression.

Stem cell culture:
Benefits and challenges

In vitro stem cell culture has emerged as a powerful tool in a variety of life science applications. Culturing adult and embryonic stem cells as well as iPSCs derived from both patient and healthy donors in special conditions allows them to be differentiated into specific cell types. Due to their source-specific origin, these cells can be used to recapitulate in vivo human cellular physiology to offer accurate, reproducible and physiologically relevant disease models.

These models are helping to uncover important biological pathways in diseases and identify novel therapeutic targets. In pre-clinical drug screening, they can also help to reliably predict the efficacy and toxicity of new drugs to accelerate drug development.

Stem cell culture also has exciting applications in gene therapy, which is an emerging but promising method of disease treatment, including for a variety of cancers. Primary stem cells (usually hematopoietic stem cells) are genetically modified via genetic engineering techniques, which involve inserting, modifying or deleting genetic material via transfection. The aim is that the genetic modification will produce a therapeutic effect when the modified cells are injected into the patient. As stem cells can self-renew, a key advantage of using them in gene therapy is that repeated administrations of the therapy can be reduced or even eliminated.

Despite the many benefits of stem cell culture, there are some challenges involved in its clinical application. For example, it can be difficult to find suitable cell culture conditions to promote proliferation but also maintain the desired stem cell properties. Moreover, the need for a high quality and quantity of cells requires large-scale expansion of stem cells, followed by efficient and homogeneous differentiation into functional derivatives. Traditional methods using two-dimensional (2D) culturing techniques (for example, using plastic culture plates and xenogenic media) provide limited expansion and lose clonal and differentiation capacity after long-term passaging. Efficient and large-scale transfection of stem cells and generation of iPSCs has also proved challenging.

A range of innovative solutions are overcoming these challenges. For example, we offer a range of specialized xeno-free growth and differentiation media for stem cell cultures that have been designed to achieve optimum cell differentiation and performance.

Our advanced electroporation method, Nucleofector^® Technology, also offers a transfection solution for hard-to-transfect stem cells. This technology enables highly efficient non-viral transfection of primary adult stem cells, genome editing in iPSCs or embryonic stem cells, and episomal reprogramming of a variety of cells into iPSCs. It also enables flexible scaling to allow you to transfect the sufficient number and type of cells for your application.

Different types of stem cells

There are two main types of stem cells used in stem cell culture: embryonic (pluripotent) and adult stem cells. Embryonic stem cells (ESCs) are obtained by extracting cells from very early embryos (blastocysts) that have been donated for research purposes. These so-called pluripotent stem cells, which are often termed ‘true’ stem cells, have the potential to differentiate into almost any cell in the body, including muscle, blood, heart and nerve cells. They are primarily involved in growth and early development.

In contrast, adult stem cells, also called tissue-specific stem cells, are stem cells that produce a limited set of specialized cells that are characteristic of a particular tissue. For example, hematopoietic stem cells in bone marrow and umbilical cord blood make all the different types of blood cells. Adult stem cells play a vital role in tissue repair due to natural wear and tear, injury or disease.

In the following sections, we outline the types of adult and pluripotent stem cells and culture systems we offer, and how they can support your applications. You can also find out how our CellBio Services can expand your research options, including custom generation of cell-based assays and modification of our existing catalog products.

Adult stem cells

We offer a range of cryopreserved adult stem cells and accompanying culture kits that are designed to meet the needs of your application. All our products are quality controlled and supplied with a certificate of analysis, as well as our recommended protocols and storage. We also recommend specialized reagents and media that have been optimized to guarantee the proliferation and performance of your stem cell cultures.

Mesenchymal stem cells (MSCs)
Hematopoietic stem cells (HSCs)

Mesenchymal stem cells

Mesenchymal stem cells (MSCs) are rare progenitor cells most commonly found in bone marrow. MSCs either divide as undifferentiated cells or differentiate into bone, cartilage, fat, muscle, tendon and marrow stroma. They are most frequently isolated from bone marrow, but they can be isolated from other tissues including adipose tissue and dental pulp.

We support our cryopreserved bone marrow-, adipose- and dental pulp-derived MSCs, with specialized growth and differentiation media and reagents for optimum cell performance. Both adipose- and dental pulp-derived stem cells demonstrate very similar phenotypic and functional characteristics to that of bone marrow-derived MSCs. All three types have been reported to be multipotent, meaning that they can differentiate down many different lineages including chondrogenic, osteogenic, adipogenic and neural.

Human mesenchymal stem cells

Our Poietics^® Normal Human Bone Marrow Derived Mesenchymal Stem Cells (hMSCs) are isolated from normal (non-diabetic) adult bone marrow withdrawn from bilateral punctures of the posterior iliac crests of normal volunteers.

See hMSC product

Adipose derived stem cells

We offer Poietics^® Normal Human Adipose Derived Stem Cells (ADSCs) derived from normal (non-diabetic) adult lipoaspirates collected during elective surgical liposuction procedures.

See hADSC product

Diabetic human adipose derived stem cells

Human Adipose Derived Stem Cells (ADSC) from Type I and Type II diabetic donors are also available as a separate product for research use: Poietics^® Diabetic Human Adipose Derived Stem Cells.

See diabetic ADSC product

Human dental pulp stem cells

Our Poietics^® Human Dental Pulp Stem Cells (DPSCs) are isolated from adult third molars collected during the extraction of a donor's 'wisdom' teeth.

See DPSC product

Hematopoietic stem cells

Hematopoiesis is the process by which blood cells are produced. Hematopoietic stem cells (HSCs) play a crucial role in hematopoiesis. HSCs, which are generally found in the bone marrow, are constantly self-renewing and differentiating into all the different types of blood cells. During this process, HSCs leave the bone marrow and enter the peripheral blood and tissues. Here, they progress through two different progenitor stages (lymphoid and myeloid progenitors) before becoming mature blood cell types:

Myeloid progenitor cells can give rise to the myeloid lineage including platelets, eosinophils, basophils, neutrophils, monocytes, and erythrocytes.
Lymphoid progenitor cells can differentiate into T-cells, B-cells and natural killer cells, which are key players in adaptive immunity.

Assessing the presence of the CD34 marker in HSCs/progenitors (a transmembrane phosphoglycoprotein protein encoded by the CD34 gene) can help to segment undifferentiated HSCs and progenitor cells.

Our broad range of HSCs with guaranteed viable cell counts and purity claims. We can obtain these from a wide range of tissue sources and cell types including bone marrow, peripheral blood immune cells and umbilical cord blood cells (all from carefully screened donors). These products can be accompanied by our recommended protocols and media, are ready to use after thawing and culturing, and are supported by an optimized culture system. This avoids having to spend time finding the right donors and isolating specific cell types, helping to simplify your research and streamline operational processes.

The types of HSC sources and cell types we offer are:

Fresh unprocessed bone marrow: We have been committed to our IRB-approved bone marrow program for over 20 years, providing the research community with fresh and viable unprocessed bone marrow to ensure relevant results, while maintaining the well-being of our donors.
Poietics^® Purified Bone Marrow Cells, including CD34+ cells, mononuclear cells, and stromal feeder cells, along with our recommended HPGMTM Hematopoietic Growth Medium for optimized proliferation during culture.
Umbilical cord blood cells: The cellular composition of umbilical cord blood is very similar to that of bone marrow. The cord blood cells we have available include purified hematopoietic stem (HSCs) and progenitor cells, lymphocytes, monocytes, and mesenchymal stem cells. We also recommend our HPGM^® Hematopoietic Growth Medium to optimize cell growth and maintenance. We have established an umbilical cord blood acquisition program to ensure you have easy access to the cord blood cells you need for your research.
Our Poietics^® Primary Human Immune Cells are isolated from peripheral blood of healthy donors. These include CD4+ cells, natural killer cells, CD14+ monocytes, peripheral blood mononuclear cells (PBMCs), and dendritic cells. These are supported by our recommended LGM-3^® Lymphocyte Growth Medium to ensure optimal cell performance.

References

Kim N and Cho SG. Clinical applications of mesenchymal stem cells Korean J intern Med 2013 Jul; 28(4):387-402