Pooled human serum: A new culture supplement for bioreactor-based cell therapies. Preliminary results

Savelli, Sara; Trombi, Luisa; D’Alessandro, Delfo; Moscato, Stefania; Pacini, Stefania; Giannotti, Stefano; Lapi, Simone; Scatena, F.; Petrini, Mario

doi:10.1016/j.jcyt.2017.12.013

Cited by 15 publications

(11 citation statements)

References 29 publications

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“…Other studies have reported the presence of a sub-population of progenitor cells in Quantum® cultures harvested after bone marrow aspirates were added directly to the bioreactor [59]. The reasons for this sub-population persisting only in Quantum® bone marrow isolations are unclear, but could be related to the lack of bone marrow density gradient centrifugation prior to seeding, or the cryoprecipitate coating of the bioreactor.…”

Section: Discussionmentioning

confidence: 99%

A comprehensive characterisation of large-scale expanded human bone marrow and umbilical cord mesenchymal stem cells

et al. 2019

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BackgroundThe manufacture of mesenchymal stem/stromal cells (MSCs) for clinical use needs to be cost effective, safe and scaled up. Current methods of expansion on tissue culture plastic are labour-intensive and involve several ‘open’ procedures. We have used the closed Quantum® hollow fibre bioreactor to expand four cultures each of MSCs derived from bone marrow (BM) and, for the first time, umbilical cords (UCs) and assessed extensive characterisation profiles for each, compared to parallel cultures grown on tissue culture plastic.MethodsBone marrow aspirate was directly loaded into the Quantum®, and cells were harvested and characterised at passage (P) 0. Bone marrow cells were re-seeded into the Quantum®, harvested and further characterised at P1. UC-MSCs were isolated enzymatically and cultured once on tissue culture plastic, before loading cells into the Quantum®, harvesting and characterising at P1. Quantum®-derived cultures were phenotyped in terms of immunoprofile, tri-lineage differentiation, response to inflammatory stimulus and telomere length, as were parallel cultures expanded on tissue culture plastic.ResultsBone marrow cell harvests from the Quantum® were 23.1 ± 16.2 × 106 in 14 ± 2 days (P0) and 131 ± 84 × 106 BM-MSCs in 13 ± 1 days (P1), whereas UC-MSC harvests from the Quantum® were 168 ± 52 × 106 UC-MSCs after 7 ± 2 days (P1). Quantum®- and tissue culture plastic-expanded cultures at P1 adhered to criteria for MSCs in terms of cell surface markers, multipotency and plastic adherence, whereas the integrins, CD29, CD49c and CD51/61, were found to be elevated on Quantum®-expanded BM-MSCs. Rapid culture expansion in the Quantum® did not cause shortened telomeres when compared to cultures on tissue culture plastic. Immunomodulatory gene expression was variable between donors but showed that all MSCs upregulated indoleamine 2, 3-dioxygenase (IDO).ConclusionsThe results presented here demonstrate that the Quantum® can be used to expand large numbers of MSCs from bone marrow and umbilical cord tissues for next-generation large-scale manufacturing, without impacting on many of the properties that are characteristic of MSCs or potentially therapeutic. Using the Quantum®, we can obtain multiple MSC doses from a single manufacturing run to treat many patients. Together, our findings support the development of cheaper cell-based treatments.Electronic supplementary materialThe online version of this article (10.1186/s13287-019-1202-4) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

A comprehensive characterisation of large-scale expanded human bone marrow and umbilical cord mesenchymal stem cells

et al. 2019

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“…In this study, hABS was also used in the isolation of MSCs from tissue and cryopreservation. Savelli et al (2018) cultured MSCs in a hollow fiber, perfused bioreactor and found that a particular population of cells, the mesodermal progenitor cells (MPCs), were enriched compared to cultures in media supplemented with FBS, where only a MSC phenotype was observed. Supplementation with human AB serum was tested in a comparative study of MSC expansion in planar and microcarrier culture at reasonable scale (2 L stirred tank systems utilizing microcarriers) (Tozetti et al, 2017).…”

Section: Cell Culture Supplementsmentioning

confidence: 99%

“…Furthermore, the shorter culture time bioreactor systems required to generate comparable cell numbers to tissue culture flasks can minimise the risk of MSC senescence and phenotypic changes due to culturing in serum (Mizukami et al, 2016). Other approaches used to increase the cell growth surface area, without increasing the footprint of the bioreactor include the use of hollow fibre bioreactors (Tozetti et al, 2017;Savelli et al, 2018) as well as fixed bed perfusion systems (Sart et al, 2014). An important feature of many scale-out systems is the ability to be able to operate them in a functionally closed manner.…”

Section: Gmp Grade Availablementioning

confidence: 99%

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

et al. 2020

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Cell therapeuticsusing cells as living drugshave made advances in many areas of medicine. One of the most clinically studied cell-based therapy products is mesenchymal stromal cells (MSCs), which have shown promising results in promoting tissue regeneration and modulating inflammation. However, MSC therapy requires large numbers of cells, the generation of which is not feasible via conventional planar tissue culture methods. Scale-up manufacturing methods (e.g., propagation on microcarriers in stirred-tank bioreactors), however, are not specifically tailored for MSC expansion. These processes may, in principle, alter the cell secretome, a vital component underlying the immunosuppressive properties and clinical effectiveness of MSCs. This review outlines our current understanding of MSC properties and immunomodulatory function, expansion in commercial manufacturing systems, and gaps in our knowledge that need to be addressed for effective up-scaling commercialization of MSC therapy.

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“…fatty acids), supplements (e.g., fetal bovine serum vs . platelet lysate), expansion protocols (e.g., multi-well plates vs. bioreactor), and cryopreservation (e.g., cryogen and freeze-thaw cycles), few studies have elucidated the influence of these changes on hMSC functional properties and clinical outcome (1, 6, 34–38).…”

Section: Hmscs Metabolic Plasticity and Culture-induced Metabolic Chamentioning

confidence: 99%

Metabolism in Human Mesenchymal Stromal Cells: A Missing Link Between hMSC Biomanufacturing and Therapy?

Yuan

Logan

2019

Front. Immunol.

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Human mesenchymal stem cells (hMSCs) are the most commonly-tested adult stem cells in cell therapy. While the initial focus for hMSC clinical applications was to exploit their multi-potentiality for cell replacement therapies, it is now apparent that hMSCs empower tissue repair primarily by secretion of immuno-regulatory and pro-regenerative factors. A growing trend in hMSC clinical trials is the use of allogenic and culture-expanded cells because they are well-characterized and can be produced in large scale from specific donors to compensate for the donor pathological condition(s). However, donor morbidity and large-scale expansion are known to alter hMSC secretory profile and reduce therapeutic potency, which are significant barriers in hMSC clinical translation. Therefore, understanding the regulatory mechanisms underpinning hMSC phenotypic and functional property is crucial for developing novel engineering protocols that maximize yield while preserving therapeutic potency. hMSC are heterogenous at the level of primary metabolism and that energy metabolism plays important roles in regulating hMSC functional properties. This review focuses on energy metabolism in regulating hMSC immunomodulatory properties and its implication in hMSC sourcing and biomanufacturing. The specific characteristics of hMSC metabolism will be discussed with a focus on hMSC metabolic plasticity and donor- and culture-induced changes in immunomodulatory properties. Potential strategies of modulating hMSC metabolism to enhance their immunomodulation and therapeutic efficacy in preclinical models will be reviewed.

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Pooled human serum: A new culture supplement for bioreactor-based cell therapies. Preliminary results

Cited by 15 publications

References 29 publications

A comprehensive characterisation of large-scale expanded human bone marrow and umbilical cord mesenchymal stem cells

A comprehensive characterisation of large-scale expanded human bone marrow and umbilical cord mesenchymal stem cells

Biological Considerations in Scaling Up Therapeutic Cell Manufacturing

Metabolism in Human Mesenchymal Stromal Cells: A Missing Link Between hMSC Biomanufacturing and Therapy?

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