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UNLOCKING OPTIMAL CONDITIONS FOR HEK293T CELL EXPANSION USING
ISCT 2024 |
HEK293T cells are a versatile cell line that can, among others, be used as producer cells for lentiviral vectors (LV) for cell and gene therapy. In particular, HEK293T can be used for LV production for Chimeric Antigen- |
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Upscaling and Neural Differentiation of Human Induced Pluripotent Stem Cell
ISCT 2024 |
The efficacy of current treatments targeting neurological diseases of the brain is affected by their inability to cross the blood brain barrier (BBB). Innovative study models to find a solution to this problem is beneficial. Human induced pluripotent stem cells (hiPSCs) are capable of indefinite cultivation and differentiation into nearly any cell type, including traditionally inaccessible cells critical for BBB function like astrocytes. hiPSCs thus can represent an effective alternative screening tool aiding in determining the efficacy of new treatments. The challenge in using this approach for screening is the need for billions of high quality cells. Obtaining high cell numbers is not impossible with traditional monolayer culturing, but laborious and inefficient. Bioreactor technology can limit the labor demand and simultaneously provide optimal growth conditions for hiPSCs. However, process development is required for the transition to such new cell culture devices, as multiple parameters can affect growth and quality. Thus, we aimed to develop methods for large scale hiPSC culture for subsequent differentiation to astrocytes in 3D. To achieve this, we investigated iPSC spheroid upscaling using the SCINUS bioreactor NG and direct differentiation of suspension-grown iPSCs towards astrocytes |
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Xeno-free cultivation of WJ-MSCs in the SCINUS bioreactor ISCT 2023 |
Wharton Jelly-derived Mesenchymal Stem Cells (WJ-MSCs) have emerged as a promising therapeutic agent for Cell Therapy due to their immunomodulatory and anti-inflammatory properties. In addition, they are not burdened by ethical issues and have minimal risk of malignant transformation [1]. WJ-MSCs are therefore great promise for treating a vast number of difficult-to-treat diseases, such as diabetes, Alzheimer’s and ALS. The current limitation is the upscaling |
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Transact or Dynabead-mediated T cell expansion in a controlled, ISCT 2023 |
Engineered Chimeric Antigen Receptor T cell (CAR-T) therapy has revolutionized the immunotherapy field, showing clinical efficacy in several haematological malignancies. Automation,
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Semi-Automated Large-Scale Expansion of Intrahepatic Cholangiocyte Organoids ISCT 2023 |
The liver is the second most required organ for transplantation purposes. Here, we have investigated the use of the SCINUS bioreactor for rapid |
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Investigating 2D Cell Culture Optimization for Large-Scale Production of IPSCs ISCT 2022 |
Large-scale culturing of induced pluripotent stem cells (iPSCs) is a prerequisite to fulfil the great potential these cells hold in regenerative medicine, drug discovery and disease modelling. For adherent culture of iPSCs, the interaction with substrate material is critical for the proper growth and maintenance of pluripotency. However, it is unclear how various parameters affect cellular growth and phenotype. In this study, we employed design-of-experiments (DoE) methodology to investigate the influence of coating, seeding density and exposure to ROCK inhibitor (ROCKi) on iPSC growth and pluripotency. |
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Preparing for cell culture scale-out: establishing parity of bioreactor- and flask-expanded mesenchymal stromal cell cultures Journal of Translational Medicine 2019 |
Cell-based therapies have the potential to become treatment options for many diseases, but efficient scale-out of these therapies has proven to be a major hurdle. Bioreactors can be used to overcome this hurdle, but changing the culture method can introduce unwanted changes to the cell product. Therefore, it is important to establish parity between products generated using traditional methods versus those generated using a bioreactor. (Open PDF) |
| A single-step expansion system for large-fold expansion of bone marrow-derived MSCs ISCT 2018 |
Cell therapies require (cost-)effective production to ensure that novel therapies are commercially viable. Closed, automated bioreactors can improve handling and safety while also reducing costs by limiting operator involvement, clean room requirements and expenditure of consumables. However, current closed solutions do not support the expansion to hundreds of millions cells from the limited initial cell numbers found in a biopsy without multiple reseeding steps. We developed novel bioreactor technology (Figure 1) with which high cell numbers can be grown from a bone marrow biopsy in a single expansion system, eliminating the need for labourand cost-intensive expansion protocols. (Open PDF) |
| Optimization of Microcarrier-based Culture of Muscle Precursor Cells ISCT 2018 |
Stress urinary incontinence (SUI) affects over 200 million people worldwide. A novel approach to treat SUI is to locally administer autologous muscle precursor cells (MPCs) into the defective sphincter muscle. For the treatment of one patient millions of cells are required. Therefore the production of these MPCs needs to be scaled up. Scinus Cell Expansion BV developed a novel bioreactor technology using a microcarrier based expansion process which provides a safe and (cost-) effective procedure for the clinical upscale of MPCs. Here, we present the optimization of a microcarrier based culture of MPCs, using a downscaled model of the SCINUS technology. (Open PDF) |
| Single-step expansion of adipose-derived stem cells with platelet lysate in SCINUS Cell expansion system ISCT 2018 |
Adipose-derived stem cells (ASCs) can be isolated from fat tissue obtained after e.g. abdominoplasty. Advantages of fat tissue over bone marrow as a source for stem cells include easier accessibility and availability of larger volumes. Costeffective production of cellular therapies requires efficient culture platforms that address major cost drivers: labor costs, clean room requirements and consumable expenditure. At the same time, process automation can increase quality and reliability of the cell product. Here we present the culture of over 500 million ASCs starting directly from a stromal vascular fraction in medium supplemented with human platelet lysate (hPL) using our SCINUS Cell Expansion system (figure 1). Human PL was used as an alternative to FBS as it contains no animal derived products and it is a rich source for varying growth factors. (Open PDF) |
| Large-scale expansion of MSCs using one-step, closed-system bioreactor technology. ISCT 2017 |
Cost-effective production of cellular therapies requires efficient culture platforms that address major cost drivers: labour costs, clean room requirements and consumable expenditure. At the same time, process automation can increase quality and reliability of the cell product. Therapies using mesenchymal stem cells (MSCs) represent a major part of cell-based clinical trials. Consequently, this cell type serves as an excellent source to demonstrate (cost-) effective culture using bioreactor technology. We demonstrate efficient large-scale culture of MSCs, using the Scinus Cell Expansion system. (Open PDF) |
| Culture of Adipose-derived Stem Cells on Microcarriers using the Scinus Cell Expansion bioreactor. ISCT 2017 |
Adipose-derived stem cells (ASCs) can be isolated from fat tissue obtained after abdominoplasty. Advantages of fat tissue over bone marrow as a source for stem cells include the easier accessibility, and availability of larger volumes. For the production of stem cells for cell therapy in patients, an upgrade to clinical large scale culture (> 200×106 cells) is necessary. Clinical scale cultures require a reproducible and efficient process. For this, a microcarriers based culture is a very suitable method. Within the Scinus Cell Expansion system (see Figure 1) adherent cells can be cultured on microcarriers in a closed environment under GMP conditions. A process for culturing large quantities of ASCs using microcarriers (MCs) using the Scinus Cell Expansion system was developed. (Open PDF) |
| Culture of adipose-derived stem cells on microcarriers. ISCT 2016 |
Adipose-derived stem cells (ASCs) can be isolated from fatty tissue. Similar to MSCs isolated from bone marrow, ASCs have multi-lineage potential and can be used as a potential source in regenerative medicine. Additionally, fat tissue is more accessible than bone marrow, and larger volumes can be obtained. For the production of cells for cell therapy in patients, an upgrade to clinical large scale culture (> 200×106 cells) is necessary. Clinical scale cultures require a reproducible and efficient process. Therefore, a process for culturing of large quantities of ASCs using microcarriers (MCs) was developed. (Open PDF) |
| One-step bone marrow-derived msc culture using novel bioreactor technology. ISCT 2016 |
Culture of bone marrow-derived MSCs for clinical application is a costly process, in large part due to the requirement for cleanroom facilities necessitated by numerous open procedures. The use of closed bioreactor systems can reduce costs and improve quality of the final cell product. However, these systems are usually ill suited to culture cells to large quantities directly from an aspirate. Here we present a closed system to culture millions of MSCs starting from a small volume bone marrow aspirate, while retaining MSC properties. (Open PDF) |
| Cost-effecient, closed system MSC culture to therapeutically relevant quantities. ISCT 2016 |
Achieving cost-efficient production of cell therapies is a major challenge, with medium costs and operator handling being significant contributors. Medium usage can be greatly reduced by using microcarrier-based expansion to reach high cell numbers in minimal volume. Microcarriers (MC) also enable closed, singlestep procedures in bioreactors that limit operator involvement and clean room requirements. Scinus Cell Expansion System is a bioreactor designed for the culture of adherent cells in a closed, single-use bag. (Open PDF) |