Characterization of human mesenchymal stem cells from multiple donors and the implications for large scale bioprocess development

Heathman, Thomas R.J.; Rafiq, Qasim A.; Chan, Alexander K.C.; Coopman, Karen; Nienow, Alvin W.; Kara, Bo; Hewitt, Christopher J.

doi:10.1016/j.bej.2015.06.018

Cited by 76 publications

(87 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Importantly however, the work also showed that the strategy adopted did not distort the main aims of the experiments. This conclusion could be reached because the relative performance of the cell lines from the different donors reported for growth in T-flasks [14] was the same during experiments with the same cells on microcarriers (data not shown). …”

Section: Overviewmentioning

confidence: 61%

Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms

Nienow

Hewitt

Heathman

et al. 2016

Biochemical Engineering Journal

Self Cite

View full text Add to dashboard Cite

Please cite this article in press as: A.W. Nienow, et al., Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms, Biochem. Eng. J. (2015), http://dx.a b s t r a c t In our recent work in different bioreactors up to 2.5 L in scale, we have successfully cultured hMSCs using the minimum agitator speed required for complete microcarrier suspension, N JS. In addition, we also reported a scaleable protocol for the detachment from microcarriers in spinner flasks of hMSCs from two donors. The essence of the protocol is the use of a short period of intense agitation in the presence of enzymes such that the cells are detached; but once detachment is achieved, the cells are smaller than the Kolmogorov scale of turbulence and hence not damaged. Here, the same approach has been effective for culture at N JS and detachment in-situ in 15 mL ambr TM bioreactors, 100 mL spinner flasks and 250 mL Dasgip bioreactors. In these experiments, cells from four different donors were used along with two types of microcarrier with and without surface coatings (two types), four different enzymes and three different growth media (with and without serum), a total of 22 different combinations. In all cases after detachment, the cells were shown to retain their desired quality attributes and were able to proliferate. This agitation strategy with respect to culture and harvest therefore offers a sound basis for a wide range of scales of operation.

show abstract

Section: Overviewmentioning

confidence: 61%

Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms

Nienow

Hewitt

Heathman

et al. 2016

Biochemical Engineering Journal

Self Cite

View full text Add to dashboard Cite

show abstract

“…adipogenesis, chondrogenesis and osteogenesis) and the secretion of a range of trophic factors that could determine specialised tissue regeneration . Similarly to other adherent cell types, hMSCs can be expanded in vitro on solid cell culture substrates such as polystyrene dishes or micrometer‐sized beads (i.e. microcarriers) .…”

Section: Introductionmentioning

confidence: 99%

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

Hanga

Murasiewicz

Pacek

et al. 2017

J. Chem. Technol. Biotechnol

Self Cite

View full text Add to dashboard Cite

BACKGROUNDHuman mesenchymal stem/stromal cells (hMSCs) are at the forefront of regenerative medicine applications due to their relatively easy isolation and availability in adults, potential to differentiate and to secrete a range of trophic factors that could determine specialised tissue regeneration. To date, hMSCs have been successfully cultured in vitro on substrates such as polystyrene dishes (TCPS) or microcarriers. However, hMSC sub‐cultivation and harvest typically employs proteolytic enzymes that act by cleaving important cell membrane proteins resulting in long‐term cell damage. In a process where the cells themselves are the product, a non‐enzymatic and non‐damaging harvesting approach is desirable.RESULTSAn alternative system for hMSC expansion and subsequent non‐enzymatic harvest was investigated here. A liquid/liquid two‐phase system was proposed, comprising a selected perfluorocarbon (FC40) and growth medium (DMEM). The cells exhibited similar cell morphologies compared with TCPS. Moreover, they retained their identity and differentiation potential post‐expansion and post‐harvest. Further, no significant difference was found when culturing hMSCs in the culture systems prepared with either fresh or recycled FC40 perfluorocarbon.CONCLUSIONSThese findings make the FC40/DMEM system an attractive alternative for traditional cell culture substrates due to their ease of cell recovery and recyclability, the latter impacting on overall process costs. © 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

show abstract

“…16 Examples of critical quality attributes (CQAs) that would benefit from sensors and enhance bioreactor performance include: small molecules and ions (pH, oxygen, and carbon dioxide), metabolites (glucose, lactate, and ammonia), and large macromolecules (enzymes, growth factors, and cytokines). [17][18][19] B. Importance of measurement and sensors in bioprocessing Quantification of essential molecules and ions has underpinned our knowledge of biological processes.…”

Section: A Emerging Cell and Gene Therapy Manufacturing Paradigmsmentioning

confidence: 99%

“…Conversely, online monitoring allows real-time quantification of analytes of interest. 17 This will yield live information during bioprocessing, which will be essential for the early detection of deviations from method parameters and enable implementation of countermeasures.…”

Section: A Emerging Cell and Gene Therapy Manufacturing Paradigmsmentioning

confidence: 99%

Enhancing cell and gene therapy manufacture through the application of advanced fluorescent optical sensors (Review)

Harrison

Chauhan

2017

Biointerphases

View full text Add to dashboard Cite

Cell and gene therapies (CGTs) are examples of future therapeutics that can be used to cure or alleviate the symptoms of disease, by repairing damaged tissue or reprogramming defective genetic information. However, despite the recent advancements in clinical trial outcomes, the path to wide-scale adoption of CGTs remains challenging, such that the emergence of a “blockbuster” therapy has so far proved elusive. Manufacturing solutions for these therapies require the application of scalable and replicable cell manufacturing techniques, which differ markedly from the existing pharmaceutical incumbent. Attempts to adopt this pharmaceutical model for CGT manufacture have largely proved unsuccessful. The most significant challenges facing CGT manufacturing are process analytical testing and quality control. These procedures would greatly benefit from improved sensory technologies that allow direct measurement of critical quality attributes, such as pH, oxygen, lactate and glucose. In turn, this would make manufacturing more robust, replicable and standardized. In this review, the present-day state and prospects of CGT manufacturing are discussed. In particular, the authors highlight the role of fluorescent optical sensors, focusing on their strengths and weaknesses, for CGT manufacture. The review concludes by discussing how the integration of CGT manufacture and fluorescent optical sensors could augment future bioprocessing approaches.

show abstract

Characterization of human mesenchymal stem cells from multiple donors and the implications for large scale bioprocess development

Cited by 76 publications

References 24 publications

Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms

Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms

Expansion of bone marrow-derived human mesenchymal stem/stromal cells (hMSCs) using a two-phase liquid/liquid system

Enhancing cell and gene therapy manufacture through the application of advanced fluorescent optical sensors (Review)

Contact Info

Product

Resources

About