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

Schubert, Susanna; Brehm, Walter; Hillmann, Aline; Burk, Janina

doi:10.1002/cyto.a.23240

Cited by 18 publications

(26 citation statements)

References 48 publications

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“…However, the biosafety of the media components in these disclosed formulations is not ensured and warrants further analysis. Moreover, some of the studies have shown differences in the growth of MSCs derived from different tissues of the same species 16,28,31 or same tissue of different species when cultured in defined SFM indicating serum-free culture conditions are tissue and species-specific and underlines the need for optimization for MSCs from different origin and/or relevant animal species 30,44 . To overcome these limitations, proprietary serum/xeno-free media, which are commercially available, have been developed as potential alternatives for in vitro clinical-grade MSCs expansion.…”

Section: Discussionmentioning

confidence: 99%

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Bhat

Viswanathan

Chandanala

et al. 2021

Sci Rep

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Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. Although therapeutic value of MSCs is now being established through many clinical trials, issues have been raised regarding their expansion as per regulatory guidelines. Fetal bovine serum usage in cell therapy poses difficulties due to its less-defined, highly variable composition and safety issues. Hence, there is a need for transition from serum-based to serum-free media (SFM). Since SFM are cell type-specific, a precise analysis of the properties of MSCs cultured in SFM is required to determine the most suitable one. Six different commercially available low serum/SFM with two different seeding densities were evaluated to explore their ability to support the growth and expansion of BM-MSCs and assess the characteristics of BM-MSCs cultured in these media. Except for one of the SFM, all other media tested supported the growth of BM-MSCs at a low seeding density. No significant differences were observed in the expression of MSC specific markers among the various media tested. In contrary, the population doubling time, cell yield, potency, colony-forming ability, differentiation potential, and immunosuppressive properties of MSCs varied with one another. We show that SFM tested supports the growth and expansion of BM-MSCs even at low seeding density and may serve as possible replacement for animal-derived serum.

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Section: Discussionmentioning

confidence: 99%

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Bhat

Viswanathan

Chandanala

et al. 2021

Sci Rep

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“…For human MSC, commercially available serum-free culture media were developed. However, when investigating the applicability of such serum-free media in equine adipose-derived MSC, we observed differences in morphology and expression of the surface marker CD90, as well as increased aggregation and spontaneous detachment of these MSC (Schubert et al, 2018). These results underlined that culture condition requirements are species-specific with regard to nutrient and growth factor supplementation.…”

Section: Introductionmentioning

confidence: 83%

Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture

Hagen

Lehmann

Aurich

et al. 2021

Front. Bioeng. Biotechnol.

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Translation of multipotent mesenchymal stromal cell (MSC)-based therapies is advancing in human and veterinary medicine. One critical issue is the in vitro culture of MSC before clinical use. Using fetal bovine serum (FBS) as supplement to the basal medium is still the gold standard for cultivation of many cell types including equine MSC. Alternatives are being explored, with substantial success using platelet lysate-supplemented media for human MSC. However, progress lags behind in the veterinary field. The aim of this study was to establish a scalable protocol for equine platelet lysate (ePL) production and to test the ePL in equine MSC culture. Whole blood was harvested into blood collection bags from 20 healthy horses. After checking sample materials for pathogen contamination, samples from 19 animals were included. Platelet concentrates were prepared using a buffy coat method. Platelets, platelet-derived growth factor BB, and transforming growth factor β1 concentrations were increased in the concentrates compared with whole blood or serum (p < 0.05), while white blood cells were reduced (p < 0.05). The concentrates were lysed using freeze/thaw cycles, which eliminated the cells while growth factor concentrations were maintained. Donor age negatively correlated with platelet and growth factor concentrations after processing (p < 0.05). Finally, all lysates were pooled and the ePL was evaluated as culture medium supplement in comparison with FBS, using adipose-derived MSC from four unrelated donor horses. MSC proliferated well in 10% FBS as well as in 10% ePL. However, using 5 or 2.5% ePL entailed highly inconsistent proliferation or loss of proliferation, with significant differences in generation times and confluencies (p < 0.05). MSC expressed the surface antigens CD90, CD44, and CD29, but CD73 and CD105 detection was low in all culture media. Adipogenic and osteogenic differentiation led to similar results in MSC from different culture media. The buffy coat method is useful to produce equine platelet concentrate with increased platelet and reduced white blood cell content in large scales. The ePL obtained supports MSC expansion similar as FBS when used at the same concentration (10%). Further investigations into equine MSC functionality in culture with ePL should follow.

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“…Thus, serum-free medium culture techniques have been followed [ 47 ] (Table 1 ). Though such an alternative has been successful to some extent in humans, further research is required in canines and equines [ 48 ]. Human MSCs under hypoxic conditions show improved immuno-modulatory effects besides, the cells have decreased differentiation capacity and carry higher expansion rate in comparison to MSCs cultured in normoxia [ 49 - 52 ].…”

Section: Mesenchymal Stem Cell Sources and Characterizationmentioning

confidence: 99%

Mesenchymal Stem Cell-Mediated Immuno-Modulatory and Anti- Inflammatory Mechanisms in Immune and Allergic Disorders

Gugjoo

Hussain

Amarpal

et al. 2020

IAD

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Background: Mesenchymal Stem Cells (MSCs) are present in almost all the tissues of the body and act as the backbone of the internal tissue homeostasis. Among their various characteristic features, immuno-modulatory and/ anti-inflammatory properties play an important role in therapeutics. Objective: The current topic focuses on the characterization and immuno-modulatory and/ antiinflammatory properties of MSCs. To present and discuss the current status of MSCs immunomodulatory properties. Methods: Available literature on MSCs properties and patents have been detailed, critically interpreted, and discussed based upon available literature. The main focus has been on their characteristic immunomodulatory and anti-inflammatory properties though some of the basic characterization markers have also been detailed. The databases searched for the literature include PubMed, Med Line, PubMed Central, Science Direct and a few other scientific databases. Results: MSCs are present in a very limited concentration in the tissues, and as such their culture expansion becomes imperative. MSCs immuno-modulatory and anti-inflammatory roles are achieved through direct cell-cell contact and / by the release of certain factors. Such properties are controlled by micro-environment upon which currently very limited control can be exerted. Besides, further insights in the xeno-protein free culture media as against the fetal bovine serum is required. Conclusions: MSCs have been well-isolated, cultured and characterized from numerous tissues of the body. The majority of the studies have shown MSCs as immuno-compromised with immunomodulatory and / or anti-inflammatory properties except some of the latest studies that have failed to achieve the desired results and thus, demand further research. Further research is required in the area to translate the results into clinical application.

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

Cited by 18 publications

References 48 publications

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Scalable Production of Equine Platelet Lysate for Multipotent Mesenchymal Stromal Cell Culture

Mesenchymal Stem Cell-Mediated Immuno-Modulatory and Anti- Inflammatory Mechanisms in Immune and Allergic Disorders

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