Publications
Preparing for cell culture scale-out: establishing parity of bioreactor- and flask-expanded mesenchymal stromal cell culturesAbstract
Background: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.
Methods:Mesenchymal stromal cells (MSCs) are cultured in parallel using either traditional culture flasks, spinner vessels or a new bioreactor system. To investigate parity between the cells obtained from different methods, har-vested cells are compared in terms of yield, phenotype and functionality.
Results:Bioreactor-based expansion yielded high cell numbers (222–510 million cells). Highest cell expansion was observed upon culture in flasks [average 5.0 population doublings (PDL)], followed by bioreactor (4.0 PDL) and spin-ner flasks (3.3 PDL). Flow cytometry confirmed MSC identity (CD73+, CD90+ and CD105+) and lack of contaminating hematopoietic cell populations. Cultured MSCs did not display genetic aberrations and no difference in differentiation and immunomodulatory capacity was observed between culture conditions. The response to IFNγ stimulation was similar for cells obtained from all culture conditions, as was the capacity to inhibit T cell proliferation.Conclusions:The new bioreactor technology can be used to culture large amounts of cells with characteristics equivalent to those cultured using traditional, flask based, methods.Keywords:Bioreactor, Mesenchymal stromal cells, Cell therapy (Open PDF)
White Papers
Efficient expansion of mesenchymal stem cells in a closed bioreactor system.

ISCT 2017
Cell therapies typically require hundreds of millions of cells for one application. For mesenchymal stem cells (MSCs), a typical dose is based on 2 million cells/kg body weight. These cells are obtained from donors, but initial cell numbers are extremely low. Therefore, these cell numbers need to be increased dramatically before they can be administered to the patient. Standard flask-based cell culture is extremely inefficient for cell therapy production. (Open PDF)
Posters
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)
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)
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 (> 200x106 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)
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)
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)
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 (> 200x106 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)
Co Publications
Dissolvable Microcarriers for hMSC and hiPSC Production and Recovery

ISSCR 2017
A new dissolvable microcarrier technology was developed that supports efficient cell production and recovery while eliminating the need for microcarrier cell separation. Dissolvable microcarriers are made from calcium cross-linked polygalacturonic acid polymers that are easily dissolved using a solution of EDTA and pectinase. To facilitate cell adhesion in serum-containing and serum-free applications, microcarriers are pre-coated with either porcine-derived denatured Collagen or Corning® Synthemax® II, a synthetic vitronectin peptide polymer. We demonstrate human mesenchymal stem cell (hMSC) and human induced pluripotent stem cell (hiPSC) growth on dissolvable microcarriers in spinner flasks and bioreactors. Upon microcarrier dissolution, nearly 100% of cells were recovered, and cells maintained their respective phenotype and differentiation capability.
Open PDF