Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part B: Modeling and Scale-up of the System

Weber, Christian; Freimark, Denise; Pörtner, Ralf; Pino-Grace, Pablo; Pöhl, Sebastian; Wallrapp, Christine; Geigle, Peter; Czermak, Peter

doi:10.1177/039139881003301103

Cited by 31 publications

(19 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…were determined by fitting model parameters to the experimental data of the fixed-bed cultivations [27]. Comparing the constants of the cultivation in the 14 cm min enhanced the yield from 5% to 76% (Figure 9, procedure a-c; Table 2).…”

Section: Cultivation Of Hmsc-tert At Different Fixed Bed Scalesmentioning

confidence: 99%

Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part A: Inoculation, Cultivation, and Cell Harvest Procedures

Weber¹,

Freimark²,

Pörtner

et al. 2010

Int J Artif Organs

Self Cite

View full text Add to dashboard Cite

Human mesenchymal stem cells (hMSC) are a promising cell source for several applications of regenerative medicine. The cells employed are either autologous or allogenic; by using stem cell lines in particular, allogenic cells enable the production of therapeutic cell implants or tissue engineered implants in stock. For these purposes, the generally small initial cell number has to be increased; this requires the use of bioreactors, which offer controlled expansion of the hMSC under GMP-conform conditions. In this study, divided into part A and B, a fixed bed bioreactor system based on non-porous borosilicate glass spheres for the expansion of hMSC, demonstrated with the model cell line hMSC-TERT, is introduced. The system offers convenient automation of the inoculation, cultivation, and harvesting procedures. Furthermore, the bioreactor has a simple design which favors its manufacturing as a disposable unit. Part A is focused on the inoculation, cultivation, and harvesting procedures. Cultivations were performed in lab scales up to a bed volume of 300 cm³. The study showed that the fixed bed system, based on 2-mm borosilicate glass spheres, as well as the inoculation, cultivation, and harvesting procedures are suitable for the expansion of hMSC with high yield and vitality.

show abstract

Section: Cultivation Of Hmsc-tert At Different Fixed Bed Scalesmentioning

confidence: 99%

Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part A: Inoculation, Cultivation, and Cell Harvest Procedures

Weber¹,

Freimark²,

Pörtner

et al. 2010

Int J Artif Organs

Self Cite

View full text Add to dashboard Cite

show abstract

“…For example for hMSC-TERT ATMP production, it is not only important to efficiently detach cells, but also to keep them viable (and undifferentiated). This gets challenging when production takes places in dynamic systems (stirred tank reactors (Cierpka et al, 2013;Elseberg et al, 2012), fixed bed-systems (Weber et al, 2010a;2010b;2010c) and the harvested cells are further processed (e.g., beadto-bead transfer, cell encapsulation (Freimark et al, 2010). Trypsin is unsuitable for hMSC-TERT detachment out of dynamic systems and for detachment of further processed cells.…”

Section: Discussionmentioning

confidence: 99%

CELL DETACHMENT BY PROLYL-SPECIFIC ENDOPEPTIDASE FROM <i>WOLFIPORIA COCOS</i>

Mika

Cierpka

Lange

et al. 2014

American Journal of Biochemistry and Biotechnology

View full text Add to dashboard Cite

As requirements for Advanced Therapy Medicinal Product (ATMP) production differ from other production processes (e.g., therapeutic protein production), cell detachment is often a crucial step for the process success. In most cases, cell detachment is done enzymatically. Although many peptidases are established in cell culture in R&D, e.g., Trypsin as gold standard, many of them seem to be unsuitable in ATMP production processes. Therefore, the present study investigated a novel endopeptidase used in food biotechnology for its applicability in ATMP processes where cell detachment is needed. The Prolyl-specific Peptidase (PsP) is of nonmammalian origin and considered as safe for humans. PsP was purified from the supernatant of the fungus Wolfiporia cocos. The isolation and purification resulted in an enzyme solution with 0.19 U mg −1 prolylspecific activity. By in silico analysis it was confirmed that attachment-promoting proteins can be cleaved by PsP in a similar amount than with Trypsin. Further the proteolytic activity was determined for PsP and Trypsin by using the same enzymatic assay. Detachment with both enzymes was compared for cells used in typical therapeutic production processes namely a mesenchymal stem cell line (hMSC-TERT) as a model for a cell therapeutic, Vero and MA104 cells used for viral therapeutic or vaccine production. The cell detachment experiments were performed with comparable enzyme activities (1.6 U mL −1). hMSC-TERT detachment was faster with PsP than with Trypsin. For Vero cells the detachment with PsP was not only faster but also more efficient. For MA104 cells the detachment rate with PsP was similar to Trypsin. For all cell types, detachment with PsP showed less influence on cell growth and metabolism compared to standard Trypsin.Thus, three cell types used in ATMP, viral therapeutics or vaccine production can be detached efficiently and gently with PsP. Therefore, PsP shows potential for cell detachment in ATMP and viral/vaccine production processes.

show abstract

“…The most promising systems are STRs and FBRs. STRs are well-characterized, scalable and controllable systems that are suitable for automation, whereas FBRs avoid the need for media replacement and also offer the opportunity to combine inoculation, expansion and harvest in one system [43][44][45].…”

Section: Cell Expansion In Bioreactorsmentioning

confidence: 99%

“…The homogeneity and scalability of FBR systems remain challenging despite intensive development work by Weber and colleagues [43,44,89,90] and by Elseberg [39]. In bed volumes of 14-300 mL, 2 mm diameter solid glass carriers [91] were successfully used to cultivate hMSC-TERT cells in Eagle's minimal essential medium (EMEM) with serum.…”

Section: Fixed-bed Reactormentioning

confidence: 99%

See 1 more Smart Citation

The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers

Elseberg¹,

Leber²,

Weidner³

et al. 2017

New Insights Into Cell Culture Technology

Self Cite

View full text Add to dashboard Cite

In the field of cell therapy, allogenic human mesenchymal stromal cells (hMSCs) are often used in clinical trials, creating a demand for cell mass production using efficient dynamic bioreactor systems. As an advanced therapy medicinal product (ATMP), such cells should meet certain special requirements, including product specifications requiring a production process compatible with good manufacturing practice (GMP). The development of processes in which the cells are the product therefore remains a significant challenge. This chapter describes the requirements at different steps in the upstream and downstream phases of such dynamic processes. Potential solutions are presented and future prospects are discussed, including the selection of media and carriers for the strictly adherent growing cells, allowing efficient cell adhesion and detachment. Strategies for dynamic cultivation in bioreactors are described in detail for fixed-bed and stirred-tank reactors based on GMP requirements and the integration of process analytical technology (PAT). Following cell harvest, separation and purification, the formulation and storage of the product are also described. Finally, the chapter covers important cell quality characteristics necessary for the approval of ATMPs.

show abstract

Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part B: Modeling and Scale-up of the System

Cited by 31 publications

References 35 publications

Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part A: Inoculation, Cultivation, and Cell Harvest Procedures

Expansion of Human Mesenchymal Stem Cells in a Fixed-Bed Bioreactor System Based on Non-Porous Glass Carrier – Part A: Inoculation, Cultivation, and Cell Harvest Procedures

CELL DETACHMENT BY PROLYL-SPECIFIC ENDOPEPTIDASE FROM <i>WOLFIPORIA COCOS</i>

The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers

Contact Info

Product

Resources

About