Serum and xeno‐free, chemically defined, no‐plate‐coating‐based culture system for mesenchymal stromal cells from the umbilical cord

Wu, Xiaoyun; Kang, Huiyan; Liu, Xuemin; Gao, Jin; Zhao, Kun; Ma, Zhi-Jie

doi:10.1111/cpr.12279

Cited by 30 publications

(34 citation statements)

References 47 publications

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“…Overall, the summary of different studies presented in Table shows that MSC expansion in serum‐free medium was repeatedly feasible and cell proliferation was often superior compared to control samples cultured in FBS‐supplemented media. However, in most studies including those on human MSCs, the immunophenotype of MSCs was not as consistent as recommended by the ISCT . However, exact percentages of cells expression positive or exclusion markers were not always given .…”

Section: Discussionmentioning

confidence: 98%

“…Finally, serum‐free culture conditions used in the published studies involved different culture media, most of which are commercially available thus their compositions are undisclosed and cannot be compared. Disclosure of serum‐free medium components is still rare in the literature, although promising results have recently been published, describing the use of a chemically defined medium for human umbilical cord MSC culture .…”

Section: Discussionmentioning

confidence: 99%

“…However, exact percentages of cells expression positive or exclusion markers were not always given . Differentiation was assessed to different extents, but overall, most serum‐free media supported maintenance of in vitro trilineage potential or even improved it compared to control medium . However, it should be acknowledged that in vitro differentiation potential alone is not predictive of MSC potency , and that serum‐free culture altered functional characteristics of MSCs in some studies .…”

Section: Discussionmentioning

confidence: 99%

“…Published studies on serum‐free culture media mainly focus on human MSCs, which were obtained from bone marrow in the majority of studies , but also from adipose tissue or birth‐associated tissues . In contrast, serum‐free culture of large animal MSCs has only been addressed in very few studies so far .…”

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confidence: 99%

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Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture

Schubert

Brehm

Hillmann³

et al. 2017

Cytometry Pt A

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For clinical applications of multipotent mesenchymal stromal cells (MSCs), serum-free culture is preferable to standardize cell products and prevent contamination with pathogens. In contrast to human MSCs, knowledge on serum-free culture of large animal MSCs is limited, despite its relevance for preclinical studies and development of veterinary cellular therapeutics. This study aimed to evaluate the suitability of a commercially available serum-free human MSC medium for culturing equine adiposederived MSCs in comparison with human adipose MSCs. Enzyme-free isolation by explant technique and expansion of equine and human cells in the serum-free medium were feasible. However, serum-free culture altered the morphology and complicated handling of equine MSCs, with cell aggregation and spontaneous detachment of multilayers, compared to culture in standard medium supplemented with fetal bovine serum. Furthermore, proliferation and the surface immunophenotype of equine cells were more variable compared to the controls and appeared to depend on the lot of the serum-free medium. Particularly the expression of CD90 was different between experimental groups (P < 0.05), with lower percentages of CD90 1 cells found in equine MSC samples cultured in serum-free medium (5.21-83.40%) compared to standard medium (86.20-99.50%). Additionally, small subpopulations expressing MSC exclusion markers such as CD14 (0.28-11.60%), CD34 (0.00-9.87%), CD45 (0.35-10.50%), or MHCII (0.00-3.67%) were found in equine samples after serum-free culture. In contrast, human samples displayed a more consistent morphology and a consistent CD29

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Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture

Schubert

Brehm

Hillmann³

et al. 2017

Cytometry Pt A

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“…Strategies under investigation to replace FBS in mammalian cell cultures include the development of a chemically defined culture medium that is free of xenogeneic components or the use of a human serum source as a protein supplement in a culture basal medium. Progress with developing chemically defined medium is being made, but a drawback of serum‐free media is that they typically have suboptimal performance and fail to support the attachment of cells to a surface or substrate unless the culture plates are precoated with factors that facilitate cellular attachment (i.e., fibronectin) . Human protein sources as replacements for FBS include autologous adult serum, pooled AB serum, umbilical cord blood serum, and platelet lysate (PL), all of which have been used intermittently in the past to promote mammalian cell cultures .…”

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confidence: 99%

Comparative analyses of industrial‐scale human platelet lysate preparations

et al. 2017

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Background Efforts are underway to eliminate fetal bovine serum (FBS) from mammalian cell cultures for clinical use. An emerging viable replacement option for FBS is human platelet lysate (PL) either as a plasma (PL-P) or serum (PL-S) based product. Study Design and Methods Nine industrial scale PL-S manufacturing runs (i.e. lots) were performed that consisted of an average volume of 24.6±2.2 liters of pooled platelet-rich-plasma (PRP) units that were obtained from apheresis donors. Manufactured lots were compared by evaluating various biochemical and functional test results. Comprehensive cytokine profiles of PL lots and product stability testing was performed. Global gene expression profiles of Mesenchymal Stromal Cells (MSCs) when cultured with PL-S or PL-P were compared with FBS. Results Electrolyte and protein levels were relatively consistent among all PL-S lots with only slight variations in glucose and calcium levels. All nine lots were as good as or better than FBS in expanding MSCs. PL-S stored at −80°C remained stable over two years. Quantitative cytokine arrays showed similarities as well as dissimilarities in the proteins present in PL-S. Greater differences in MSC gene expression profiles were attributable to the starting cell source than as to whether PL or FBS were used as culture supplements. Conclusion Using a large-scale standardized method, lot-to-lot variations were noted for industrial scale preparations of PL-S. However, all lots were as good as or better than FBS in supporting MSC growth. Together these data indicate that off-the-shelf PL is a feasible substitute for FBS in MSC cultures.

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Identification of critical process parameters for expansion of clinical grade human Wharton's jelly‐derived mesenchymal stromal cells in stirred‐tank bioreactors

López‐Fernández,

Garcia‐Gragera,

Lecina

et al. 2024

Biotechnology Journal

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Cell therapies based on multipotent mesenchymal stromal cells (MSCs) are traditionally produced using 2D culture systems and platelet lysate‐ or serum‐containing media (SCM). Although cost‐effective for single‐dose autologous treatments, this approach is not suitable for larger scale manufacturing (e.g., multiple‐dose autologous or allogeneic therapies with banked MSCs); automated, scalable and Good Manufacturing Practices (GMP)‐compliant platforms are urgently needed. The feasibility of transitioning was evaluated from an established Wharton's jelly MSCs (WJ‐MSCs) 2D production strategy to a new one with stirred‐tank bioreactors (STRs). Experimental conditions included four GMP‐compliant xeno‐ and serum‐free media (XSFM) screened in 2D conditions and two GMP‐grade microcarriers assessed in 0.25 L‐STRs using SCM. From the screening, a XSFM was selected and compared against SCM using the best‐performing microcarrier. It was observed that SCM outperformed the 2D‐selected medium in STRs, reinforcing the importance of 2D‐to‐3D transition studies before translation into clinical production settings. It was also found that attachment efficiency and microcarrier colonization were essential to attain higher fold expansions, and were therefore defined as critical process parameters. Nevertheless, WJ‐MSCs were readily expanded in STRs with both media, preserving critical quality attributes in terms of identity, viability and differentiation potency, and yielding up to 1.47 × 109 cells in a real‐scale 2.4‐L batch.

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Serum and xeno‐free, chemically defined, no‐plate‐coating‐based culture system for mesenchymal stromal cells from the umbilical cord

Cited by 30 publications

References 47 publications

Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture

Serum‐free human MSC medium supports consistency in human but not in equine adipose‐derived multipotent mesenchymal stromal cell culture

Comparative analyses of industrial‐scale human platelet lysate preparations

Identification of critical process parameters for expansion of clinical grade human Wharton's jelly‐derived mesenchymal stromal cells in stirred‐tank bioreactors

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