Alterations of mesenchymal stromal cells in cerebrospinal fluid: insights from transcriptomics and an ALS clinical trial

Abstract Background Mesenchymal stromal cells (MSCs) have been studied with increasing intensity as clinicians and researchers strive to understand the ability of MSCs to modulate disease progression and promote tissue regeneration. As MSCs are used for diverse applications, it is important to appreciate how specific physiological environments may stimulate changes that alter the phenotype of the cells. One need for neuroregenerative applications is to characterize the spectrum of MSC responses to the cerebrospinal fluid (CSF) environment after their injection into the intrathecal space. Mechanistic understanding of cellular biology in response to the CSF environment may predict the ability of MSCs to promote injury repair or provide neuroprotection in neurodegenerative diseases. Methods In this study, we characterized changes in morphology, metabolism, and gene expression occurring in human adipose-derived MSCs cultured in human (hCSF) or artificial CSF (aCSF) as well as examined relevant protein levels in the CSF of subjects treated with MSCs for amyotrophic lateral sclerosis (ALS). Results Our results demonstrated that, under intrathecal-like conditions, MSCs retained their morphology, though they became quiescent. Large-scale transcriptomic analysis of MSCs revealed a distinct gene expression profile for cells cultured in aCSF. The aCSF culture environment induced expression of genes related to angiogenesis and immunomodulation. In addition, MSCs in aCSF expressed genes encoding nutritional growth factors to expression levels at or above those of control cells. Furthermore, we observed a dose-dependent increase in growth factors and immunomodulatory cytokines in CSF from subjects with ALS treated intrathecally with autologous MSCs. Conclusions Overall, our results suggest that MSCs injected into the intrathecal space in ongoing clinical trials remain viable and may provide a therapeutic benefit to patients.

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Seeding decellularized nerve allografts with adipose-derived mesenchymal stromal cells: An in vitro analysis of the gene expression and growth factors produced

Journal of Plastic Reconstructive & Aesthetic Surgery ◽

10.1016/j.bjps.2019.04.014 ◽

2019 ◽

Vol 72 (8) ◽

pp. 1316-1325 ◽

Cited By ~ 7

Author(s):

Nadia Rbia ◽

Liselotte F. Bulstra ◽

Eric A. Lewallen ◽

Steven E.R. Hovius ◽

Andre J. van Wijnen ◽

...

Keyword(s):

Gene Expression ◽

Growth Factors ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Vitro Analysis ◽

In Vitro Analysis

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Interleukin-1β Induced Matrix Metalloproteinase Expression in Human Periodontal Ligament-Derived Mesenchymal Stromal Cells under In Vitro Simulated Static Orthodontic Forces

International Journal of Molecular Sciences ◽

10.3390/ijms22031027 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1027

Author(s):

Christian Behm ◽

Michael Nemec ◽

Alice Blufstein ◽

Maria Schubert ◽

Xiaohui Rausch-Fan ◽

...

Keyword(s):

Gene Expression ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Periodontal Ligament ◽

Induced Expression ◽

Gene And Protein Expression ◽

Tensile Strains ◽

Orthodontic Forces ◽

Low Magnitude

The periodontal ligament (PDL) responds to applied orthodontic forces by extracellular matrix (ECM) remodeling, in which human periodontal ligament-derived mesenchymal stromal cells (hPDL-MSCs) are largely involved by producing matrix metalloproteinases (MMPs) and their local inhibitors (TIMPs). Apart from orthodontic forces, the synthesis of MMPs and TIMPs is influenced by the aseptic inflammation occurring during orthodontic treatment. Interleukin (IL)-1β is one of the most abundant inflammatory mediators in this process and crucially affects the expression of MMPs and TIMPs in the presence of cyclic low-magnitude orthodontic tensile forces. In this study we aimed to investigate, for the first time, how IL-1β induced expression of MMPs, TIMPs and how IL-1β in hPDL-MSCs was changed after applying in vitro low-magnitude orthodontic tensile strains in a static application mode. Hence, primary hPDL-MSCs were stimulated with IL-1β in combination with static tensile strains (STS) with 6% elongation. After 6- and 24 h, MMP-1, MMP-2, TIMP-1 and IL-1β expression levels were measured. STS alone had no influence on the basal expression of investigated target genes, whereas IL-1β caused increased expression of these genes. In combination, they increased the gene and protein expression of MMP-1 and the gene expression of MMP-2 after 24 h. After 6 h, STS reduced IL-1β-induced MMP-1 synthesis and MMP-2 gene expression. IL-1β-induced TIMP-1 gene expression was decreased by STS after 6- and 24-h. At both time points, the IL-1β-induced gene expression of IL-1β was increased. Additionally, this study showed that fetal bovine serum (FBS) caused an overall suppression of IL-1β-induced expression of MMP-1, MMP-2 and TIMP-1. Further, it caused lower or opposite effects of STS on IL-1β-induced expression. These observations suggest that low-magnitude orthodontic tensile strains may favor a more inflammatory and destructive response of hPDL-MSCs when using a static application form and that this response is highly influenced by the presence of FBS in vitro.

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Alteration of Hypoxia-Associated Gene Expression in Replicatively Senescent Mesenchymal Stromal Cells under Physiological Oxygen Level

Biochemistry (Moscow) ◽

10.1134/s0006297919030088 ◽

2019 ◽

Vol 84 (3) ◽

pp. 263-271 ◽

Cited By ~ 3

Author(s):

A. Yu. Ratushnyy ◽

Yu. V. Rudimova ◽

L. B. Buravkova

Keyword(s):

Gene Expression ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Oxygen Level

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Cultured Human Adipose Tissue Pericytes and Mesenchymal Stromal Cells Display a Very Similar Gene Expression Profile

Stem Cells and Development ◽

10.1089/scd.2015.0153 ◽

2015 ◽

Vol 24 (23) ◽

pp. 2822-2840 ◽

Cited By ~ 21

Author(s):

Lindolfo da Silva Meirelles ◽

Tathiane Maistro Malta ◽

Virgínia Mara de Deus Wagatsuma ◽

Patrícia Viana Bonini Palma ◽

Amélia Goes Araújo ◽

...

Keyword(s):

Gene Expression ◽

Adipose Tissue ◽

Gene Expression Profile ◽

Mesenchymal Stromal Cells ◽

Expression Profile ◽

Stromal Cells ◽

Human Adipose Tissue ◽

Similar Gene Expression Profile ◽

Similar Gene

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The effect of culture media on large-scale expansion and characteristic of adipose tissue-derived mesenchymal stromal cells

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1331-9 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 8

Author(s):

Justyna Czapla ◽

Sybilla Matuszczak ◽

Klaudia Kulik ◽

Ewa Wiśniewska ◽

Ewelina Pilny ◽

...

Keyword(s):

Adipose Tissue ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Large Scale ◽

Culture Media ◽

Scale Expansion

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Preconditioning of mesenchymal stromal cells with low-intensity ultrasound: influence on chondrogenesis and directed SOX9 signaling pathways

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1532-2 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Neety Sahu ◽

Gaurav Budhiraja ◽

Anuradha Subramanian

Keyword(s):

Gene Expression ◽

Protein Expression ◽

Signaling Pathways ◽

Mesenchymal Stromal Cells ◽

Nuclear Localization ◽

Stromal Cells ◽

Actin Reorganization ◽

Low Intensity ◽

Gene And Protein Expression ◽

Low Intensity Ultrasound

Abstract Background Continuous low-intensity ultrasound (cLIUS) facilitates the chondrogenic differentiation of human mesenchymal stromal cells (MSCs) in the absence of exogenously added transforming growth factor-beta (TGFβ) by upregulating the expression of transcription factor SOX9, a master regulator of chondrogenesis. The present study evaluated the molecular events associated with the signaling pathways impacting SOX9 gene and protein expression under cLIUS. Methods Human bone marrow-derived MSCs were exposed to cLIUS stimulation at 14 kPa (5 MHz, 2.5 Vpp) for 5 min. The gene and protein expression of SOX9 was evaluated. The specificity of SOX9 upregulation under cLIUS was determined by treating the MSCs with small molecule inhibitors of select signaling molecules, followed by cLIUS treatment. Signaling events regulating SOX9 expression under cLIUS were analyzed by gene expression, immunofluorescence staining, and western blotting. Results cLIUS upregulated the gene expression of SOX9 and enhanced the nuclear localization of SOX9 protein when compared to non-cLIUS-stimulated control. cLIUS was noted to enhance the phosphorylation of the signaling molecule ERK1/2. Inhibition of MEK/ERK1/2 by PD98059 resulted in the effective abrogation of cLIUS-induced SOX9 expression, indicating that cLIUS-induced SOX9 upregulation was dependent on the phosphorylation of ERK1/2. Inhibition of integrin and TRPV4, the upstream cell-surface effectors of ERK1/2, did not inhibit the phosphorylation of ERK1/2 and therefore did not abrogate cLIUS-induced SOX9 expression, thereby suggesting the involvement of other mechanoreceptors. Consequently, the effect of cLIUS on the actin cytoskeleton, a mechanosensitive receptor regulating SOX9, was evaluated. Diffused and disrupted actin fibers observed in MSCs under cLIUS closely resembled actin disruption by treatment with cytoskeletal drug Y27632, which is known to increase the gene expression of SOX9. The upregulation of SOX9 under cLIUS was, therefore, related to cLIUS-induced actin reorganization. SOX9 upregulation induced by actin reorganization was also found to be dependent on the phosphorylation of ERK1/2. Conclusions Collectively, preconditioning of MSCs by cLIUS resulted in the nuclear localization of SOX9, phosphorylation of ERK1/2 and disruption of actin filaments, and the expression of SOX9 was dependent on the phosphorylation of ERK1/2 under cLIUS.

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