Cell Therapy
From basic discoveries to preclinical solutions

Expedite research success from basic discoveries to preclinical solutions. OLS, Your Partner in Cell Research, supports this endeavour. OLS accompanies in cell therapy research with high-quality devices, cell culture consumables and individualized services tailored to any specific needs. This entails reliable support at all stages, from basic in vitro research to preclinical in vivo studies with the final goal to find a cure for certain diseases.
Cell therapy is an umbrella term for several categories such as (i) Immunotherapy (e.g., CAR-T-cell therapy), (ii) Stem cell therapy, (iii) Bone marrow transplantation and (iv) Blood transfusion.
The final goal of cell therapy is to inject or transplant (allogeneic or autologous) cells into a patient to restore the lost function of an organ or to fight certain diseases. In the case of CAR-T-cell therapy, e.g. cancer cells are meant to be destroyed by taking advantage of the patient´s own immune system.

Figure 1: Electron microscope image showing T-cells, pseudo-colored in pink, interacting with dendritic cells, pseudo-colored in grey, which is an essential process taking place, for example, in vaccine development. Antigen-presenting dendritic cells have been reported to be involved in boosting the tumor-killing effect by CAR-T cells. Picture courtesy: Texas Center for Cancer Nanomedicine (TCCN).

To enhance the entirety of the cell therapy development process, OLS provides multiple innovative technologies. This includes technologies that are able to measure the cytotoxic effect of T-cells engineered in a laboratory to kill cancer cells. With these devices – Live Cell Analysis System xCELLigence – it is also possible to monitor the efficacy of manipulated T-cells meant to be used for chimeric antigen receptor T (CAR-T) cell therapy.

Figure 2. Electron microscopy image showing erythrocytes and leukocytes. The receptors of T-lymphocytes, a subfraction of the leukocytes, can be manipulated resulting in CAR-T cells. Picture courtesy: National Cancer Institute, USA.

Flow Cytometry NovoCyte Instruments in combination with the vast portfolio of Proteintech antibodies directed against CD markers Antibodies for Flow Cytometry, are the ideal tools for the identification of subpopulations of blood cells (e.g., T-cells for CAR-T-cell therapy) or for developing new cell lines.
Enhance the reproducibility of T-cell experiments with the complete, serum-free, xeno-free T-cell expansion medium smarT™.

Induced pluripotent stem cells (iPSC) can be differentiated into almost any cell type and in this differentiated form they can be injected (allogeneic or autologous) into animals or patients to restore the function of tissues. MSC (mesenchymal stem cells, mesenchymal stromal cells) can be differentiated into chondrocytes, adipocytes and osteoblasts with the final goal to substitute cartilage, adipose or bone tissue.
To optimize the quality control, expansion, differentiation and storage of your iPSC and MSC, a multitude of options are available:
• 3D Cell Culture Bioreactor CERO 3D
• Karyotyping Teaching Kits as part of our HiMedia catalog
• Dissociation reagents, such as Accutase™ Cell Culture Reagents
• CryoXL™ Freezing solution
• HiMesoXL™ and STEMin 1™ mesenchymal stem cell media as part of our HiMedia catalog

For clonal selection, several iPSC clones can be dispensed into 96 well plates assuring high expression of pluripotency markers with our Hana https://www.namocell.com/hana-single-cell-dispenser and Pala Single Cell Dispenser – Namocell.

Bone marrow transplantation, blood transfusion and hematopoietic stem cells (HSC)

NovoCyte Flow Cytometry Instruments Flow Cytometry NovoCyte Instruments in combination with Proteintech antibodies directed against surface markers Antibodies for Flow Cytometry are perfect tools to identify certain cell types from bone marrow and blood samples. HiMarrowXL™ karyotyping medium is suitable for in vitro cultures of bone marrow cells assuring a high mitotic index, ideal chromosome resolution and is delivered in convenient 10 ml bottles to avoid repeated freeze-thaw cycles. HiSep™ Density Gradient Separation Media and Ficoll will prove instrumental in separating peripheral blood mononuclear cells (PBMC). IMDM (Iscove’s modified Dulbecco’s medium) is highly enriched (e.g., in glucose, vitamins and amino acids) and optimized for cultivating HSC.
Quality Control of cells is crucial, particularly if the cells are meant to be used for injection into patients, so it is important to cover quality control with the most reliable and precise tools:
• CASY Cell Counter & Analyzer (OLS OMNI Life Science)
• xCELLigence
• zenCELL owl
• Mycoplasma detection and elimination kits as part of our HiMedia catalog

If quality checks by these measures are carried out on a regular basis, corrective action and preventive action (CAPA) can be applied which improves quality, saves time, work and costs.

CAR-T cells are promising candidates for immunotherapy of cancer cells. Their therapeutic potential is based on chimeric antigen receptors (CAR) presented on the T-cell surface and optimized to recognize cancer-specific antigens on tumor cell surfaces to efficiently induce tumor cell killing (Fig. 3).

Figure 3: Scheme of the CAR-T cell therapy

The easy way to investigate the potency of new CAR-T constructs: xCELLigence RTCA

To screen the cytolytic potency of newly designed CAR-T cells, xCELLigence RTCA systems take advantage of different adhesion properties of target and effector cells Live Cell Analysis System xCELLigence. Adherent target cells strongly interact with gold electrodes integrated into the well bottom. Proliferation of adherent target cells will lead to increasing Cell Index values. In contrast, effector cells remain in suspension and do not interact with gold electrodes, thus do not directly affect Cell Index values (Fig. 4).

Figure 4: CAR-T cells killing target cells measured by the xCELLigence RTCA

CAR-T cells, added to the target cells, recognize and bind to antigens that are presented by the target cells. Cytolytic activity of CAR-T cells is accompanied by the detachment of target cells, resulting in a decrease of Cell Index values depending on the specificity and cytolytic potency of the effector cells (Fig. 4).

The following schematic representation shows target cell killing by different CAR-T cell constructs in real time (Fig.5). Target cells were seeded into an E-Plate and analyzed continuously for a certain time. Increasing Cell Index values represent target cell proliferation. Upon addition of different CAR-T cell constructs Cell Index values started to change. Compared to “normal“ T cells, all T cells harbouring a chimeric antigen receptor (CAR-T cells) showed higher cytolytic potential, represented by a decline of Cell Index values. The reduction of Cell Index values was dependent on specificity and cytolytic activity of the effector cells. Most efficient killing was conducted by CAR-T 3. Here, Cell Index values were reduced to almost zero, indicating that most target cells were killed.

Figure 5: Efficacy of different CAR-T constructs measured by the xCELLigence RTCA

Measuring T cell-mediated cytolysis of target cells is just one great example of how the xCELLigence technology can boost your research. xCELLigence RTCA systems can also be used for a wide variety of further approaches, like cell-proliferation, cytotoxicity, invasion/migration, barrier function, virology and cardio toxicity. The xCELLigence family Live Cell Analysis System xCELLigence comprises several devices for different throughput needs (1 x 16 wells up to 4 x 384 wells) and impedance measurements combined with live cell imaging, the state of the art xCELLigence eSight system Real-Time Live-Cell Imaging – CELLigence® RTCA eSight .”