Dynamic and social behaviors of human pluripotent stem cells

Phadnis, Smruti M.; Loewke, Nathan O.; Dimov, Ivan K.; Pai, Sunil; Amwake, Christine E.; Solgaard, Olav; Baer, Thomas M.; Chen, Bertha; Pera, Renee A. Reijo

doi:10.1038/srep14209

Cited by 18 publications

(25 citation statements)

References 21 publications

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“…These approaches could also provide insight into the progression and establishment of MSC heterogeneity as it relates to differences in donor (or tissue) source and in manufacturing (40,53). Advancements in live-cell tracking and monitoring in vitro would further allow for direct correlation of early morphological phenotypes with assay outcomes (15,54).…”

Section: Morphological Features Of Mscs After Ifn-γ Stimulation Corrementioning

confidence: 99%

Morphological features of IFN-γ–stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

Klinker

Marklein

Surdo

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

168

166

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Human mesenchymal stromal cell (MSC) lines can vary significantly in their functional characteristics, and the effectiveness of MSCbased therapeutics may be realized by finding predictive features associated with MSC function. To identify features associated with immunosuppressive capacity in MSCs, we developed a robust in vitro assay that uses principal-component analysis to integrate multidimensional flow cytometry data into a single measurement of MSC-mediated inhibition of T-cell activation. We used this assay to correlate single-cell morphological data with overall immunosuppressive capacity in a cohort of MSC lines derived from different donors and manufacturing conditions. MSC morphology after IFN-γ stimulation significantly correlated with immunosuppressive capacity and accurately predicted the immunosuppressive capacity of MSC lines in a validation cohort. IFN-γ enhanced the immunosuppressive capacity of all MSC lines, and morphology predicted the magnitude of IFN-γ-enhanced immunosuppressive activity. Together, these data identify MSC morphology as a predictive feature of MSC immunosuppressive function.H uman mesenchymal stromal cells (MSCs) can potently suppress immune responses in vitro and in animal models of human disease (1, 2), but to date MSC-based therapies have produced mixed results in clinical trials for treatment of inflammatory and autoimmune diseases (3, 4). A major challenge in the development of consistently effective MSC-based immunosuppressive therapies is that MSC lines derived from different donors and manufacturing processes (i.e., cell expansion) can possess markedly dissimilar immunosuppressive function (3,5,6). Although methods exist to assess MSC immunosuppression in vitro, they are often based on only a few measured outcomes, assay culture conditions, and donor MSC samples (5, 7-9). To improve upon these methods, we developed an experimental and analytical approach to quantify MSC-mediated immune suppression using principal-component analysis (PCA) to integrate multiple measurements of T-cell activation assessed at a range of MSC densities. This approach allowed us to determine a single value for immunosuppressive capacity for MSC lines derived from two different manufacturing processes and 13 independent donors.Another major challenge associated with MSC-based immune therapies is the lack of well-defined predictive markers to identify MSC lines with therapeutically relevant biological activities or manufacturing processes that produce more effective MSCbased products. Efforts have been made to identify MSC quality attributes associated with immunosuppression (6, 7), but the majority of clinical studies (10) rely upon the surface markers described by Dominici et al. (11). Having previously shown that morphology can predict MSC mineralization capacity (12), we hypothesized that morphological features associated with immunosuppression in MSCs could be identified and used to predict their performance in our quantitative immunosuppression assay.Using our quantitative method for as...

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Section: Morphological Features Of Mscs After Ifn-γ Stimulation Corrementioning

confidence: 99%

Morphological features of IFN-γ–stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

Klinker

Marklein

Surdo

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

168

166

View full text Add to dashboard Cite

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“…Often there is still a connection between the cell bodies at these distances, but the cells move in independent directions whilst maintaining the connection, and as an isotropic random walk when considered as a whole entity [33]. Neighbouring cells are integral to colony formation as cell survival and cell divisions are highly correlated with the number of neighbouring cells [38].…”

Section: Discussionmentioning

confidence: 99%

Correlated random walks of human embryonic stem cells in vitro

et al. 2018

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We perform a detailed analysis of the migratory motion of human embryonic stem cells in two-dimensions, both when isolated and in close proximity to another cell, recorded with time-lapse microscopic imaging. We show that isolated cells tend to perform an unusual locally anisotropic walk, moving backwards and forwards along a preferred local direction correlated over a timescale of around 50 min and aligned with the axis of the cell elongation. Increasing elongation of the cell shape is associated with increased instantaneous migration speed. We also show that two cells in close proximity tend to move in the same direction, with the average separation of [Formula: see text]m or less and the correlation length of around 25 μm, a typical cell diameter. These results can be used as a basis for the mathematical modelling of the formation of clonal hESC colonies.

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“…In monolayer culture, hPSCs are typically cultured using ECM proteins such as fibronectin and lamins in addition to soluble factors such as growth factors and cytokines . The hPSCs adhere to the culture surface following single cell seeding and divide or migrate to form small colonies, which then grow and fuse . The ability of cells to divide and migrate, as single cells or as a collective, alters the spatial distribution of cells, whether it is loose or dense ECM or even other cells.…”

Section: Bioprocess Design Considerations To Enhance Scalable Expansimentioning

confidence: 99%

“…Spatial distribution of cells in monolayer culture is often observed . The different spatial distribution in culture vessel can affect growth kinetics.…”

Section: Bioprocess Design Considerations To Enhance Scalable Expansimentioning

confidence: 99%

Bioengineering Considerations for a Nurturing Way to Enhance Scalable Expansion of Human Pluripotent Stem Cells

Kim

Kino‐oka

2020

Biotechnology Journal

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Understanding how defects in mechanotransduction affect cell‐to‐cell variability will add to the fundamental knowledge of human pluripotent stem cell (hPSC) culture, and may suggest new approaches for achieving a robust, reproducible, and scalable process that result in consistent product quality and yields. Here, the current state of the understanding of the fundamental mechanisms that govern the growth kinetics of hPSCs between static and dynamic cultures is reviewed, the factors causing fluctuations are identified, and culture strategies that might eliminate or minimize the occurrence of cell‐to‐cell variability arising from these fluctuations are discussed. The existing challenges in the development of hPSC expansion methods for enabling the transition from process development to large‐scale production are addressed, a mandatory step for industrial and clinical applications of hPSCs.

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Dynamic and social behaviors of human pluripotent stem cells

Cited by 18 publications

References 21 publications

Morphological features of IFN-γ–stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

Morphological features of IFN-γ–stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

Correlated random walks of human embryonic stem cells in vitro

Bioengineering Considerations for a Nurturing Way to Enhance Scalable Expansion of Human Pluripotent Stem Cells

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