scholarly journals A conditionally immortalized Gli1-positive kidney mesenchymal cell line models myofibroblast transition

2019 ◽  
Vol 316 (1) ◽  
pp. F63-F75 ◽  
Author(s):  
Eoghainín Ó hAinmhire ◽  
Haojia Wu ◽  
Yoshiharu Muto ◽  
Erinn L. Donnelly ◽  
Flavia G. Machado ◽  
...  
Keyword(s):  
Stem Cell ◽  
Growth Factor ◽  
Mesenchymal Stem Cell ◽  
Cell Line ◽  
Transforming Growth Factor ◽  
Serum Response Factor ◽  
Primary Cultures ◽  
Kidney Fibrosis

Glioma-associated oncogene homolog-1 (Gli1)-positive resident mesenchymal stem cell-like cells are the predominant source of kidney myofibroblasts in fibrosis, but investigating Gli1-positive myofibroblast progenitor activation is hampered by the difficulty of isolating and propagating primary cultures of these cells. Using a genetic strategy with positive and negative selection, we isolated Kidney-Gli1 (KGli1) cells that maintain expression of appropriate mesenchymal stem cell-like cell markers, respond to hedgehog pathway activation, and display robust myofibroblast differentiation upon treatment with transforming growth factor-β (TGF-β). Coculture of KGli1 cells with endothelium stabilizes capillary formation. Single-cell RNA sequencing (scRNA-seq) analysis during differentiation identified autocrine ligand-receptor pair upregulation and a strong focal adhesion pathway signal. This led us to test the serum response factor inhibitor CCG-203971 that potently inhibited TGF-β-induced pericyte-to-myofibroblast transition. scRNA-seq also identified the unexpected upregulation of nerve growth factor (NGF), which we confirmed in two mouse kidney fibrosis models. The Ngf receptor Ntrk1 is expressed in tubular epithelium in vivo, suggesting a novel interstitial-to-tubule paracrine signaling axis. Thus, KGli1 cells accurately model myofibroblast activation in vitro, and the development of this cell line provides a new tool to study resident mesenchymal stem cell-like progenitors in health and disease.

Effects of Hydrostatic Pressure and Transforming Growth Factor-β3 on Adult Human Mesenchymal Stem Cell Chondrogenesis In Vitro

2006 ◽  
Vol 12 (6) ◽  
pp. 1419-1428 ◽  
Author(s):  
Keita Miyanishi ◽  
Michael C.D. Trindade ◽  
Derek P. Lindsey ◽  
Gary S. Beaupré ◽  
Dennis R. Carter ◽  
...  
Keyword(s):  
Stem Cell ◽  
Growth Factor ◽  
Hydrostatic Pressure ◽  
Mesenchymal Stem Cell ◽  
Transforming Growth Factor ◽  
Human Mesenchymal Stem Cell ◽  
Adult Human

Effects of Hydrostatic Pressure and Transforming Growth Factor-3 on Adult Human Mesenchymal Stem Cell Chondrogenesis In Vitro

2006 ◽  
Vol 0 (0) ◽  
pp. 060706073730012
Author(s):  
Keita Miyanishi ◽  
Michael C.D. Trindade ◽  
Derek P. Lindsey ◽  
Gary S. Beaupre ◽  
Dennis R. Carter ◽  
...  
Keyword(s):  
Stem Cell ◽  
Growth Factor ◽  
Hydrostatic Pressure ◽  
Mesenchymal Stem Cell ◽  
Transforming Growth Factor ◽  
Human Mesenchymal Stem Cell ◽  
Adult Human

Mesenchymal Stem Cell-Derived Vesicles Reverse Hematopoietic Radiation Damage

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2459-2459 ◽  
Author(s):  
Sicheng Wen ◽  
Laura R Goldberg ◽  
Mark S Dooner ◽  
John L Reagan ◽  
Peter J Quesenberry
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cell Line ◽  
Radiation Damage ◽  
Cell Fate ◽  
Conflicts Of Interest ◽  
Marrow Cell ◽  
Whole Body

Abstract Extracellular vesicles, including exosomes and microvesicles, have been shown to epigenetically modulate cell fate (Aliotta et al JEV, 2013). We have evaluated the effect of murine or human mesenchymal stem cell (MSC)-derived vesicles on marrow from irradiated mice or on the marrow cell line, FDC-P1, irradiated in vitro. B6.SJL mice were exposed to 0 or 100 cGy whole body irradiation and 7 days later marrow was harvested, lineage depleted and then cultured with or without the addition of murine MSC-derived vesicles for 48 hours. The cells were then engrafted into 200 cGy exposed C57BL/6J mice and engraftment was analyzed at 3, 12, 24 and 36 weeks post-transplant. Vesicle exposure led to a statistically significant increase in engraftment by the irradiated cells at 24 and 36 weeks with the average percent donor engraftment equal to 8.8 ± 4% and 9.6 ± 5% for the vesicle treated group compared to 3.7 ± 2% and 3 ± 2% for the non-vesicle treated groups at 24 weeks and 36 weeks, respectively (Figure 1). In a similar experiment, murine marrow was harvested 24 hours after 100 cGy whole body irradiation, and cultured with and without vesicles for 24 hours. There was a statistically significant increase in donor engraftment by the Lineage negative cells incubated with MSC-derived vesicles at 3 and 6 months (Figure 2). The increase in engraftment potential with vesicle exposure persisted in serial transplantation. These studies indicate that MSC-derived vesicles can reverse radiation damage to marrow when administered either 1 or 7 days after irradiation. The in vivo effect of murine MSC-derived vesicles on post-radiation blood cell count recovery was tested. C57BL/6 mice were exposed to 500 cGy total body irradiation. MSC-derived vesicles were then injected intravenously 6, 24 and 72 hours after irradiation. WBC recovery compared to non- irradiated mice was 17% in the non-vesicle group and 37% in the vesicle- injected group at 14 days; the values at 28 days were 61 and 89%, respectively, indicating that vesicle injection enhanced leukocyte recovery following radiation damage. Marrow cells from the non-irradiated, the irradiated and the irradiated plus vesicle-treated mice were then analyzed for gene expression. Twelve genes were elevated in the irradiated cells and the elevations were reversed by vesicle exposure while 17 genes were depressed in the irradiated cells and the depressions reversed by vesicle exposure. Studies with the marrow cell line, FDC-P1, showed that apoptosis in irradiated cells was reversed by exposure to MSC-derived vesicles and growth of irradiated cells was restored. Restoration of growth by in vitro exposure to MSC-derived vesicles is shown in Figure 3. In summary, these data indicate that MSC-derived vesicles have the capacity to ameliorate radiation-induced damage to marrow and may represent an important biologic for use in radiation exposures. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Uptake and Distribution of Administered Bone Marrow Mesenchymal Stem Cell Extracellular Vesicles in Retina

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 730
Author(s):  
Biji Mathew ◽  
Leianne A. Torres ◽  
Lorea Gamboa Gamboa Acha ◽  
Sophie Tran ◽  
Alice Liu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Extracellular Vesicles ◽  
Time Course ◽  
Ganglion Cells ◽  
Ex Vivo ◽  
Dependent Manner ◽  
Paracrine Effects

Cell replacement therapy using mesenchymal (MSC) and other stem cells has been evaluated for diabetic retinopathy and glaucoma. This approach has significant limitations, including few cells integrated, aberrant growth, and surgical complications. Mesenchymal Stem Cell Exosomes/Extracellular Vesicles (MSC EVs), which include exosomes and microvesicles, are an emerging alternative, promoting immunomodulation, repair, and regeneration by mediating MSC’s paracrine effects. For the clinical translation of EV therapy, it is important to determine the cellular destination and time course of EV uptake in the retina following administration. Here, we tested the cellular fate of EVs using in vivo rat retinas, ex vivo retinal explant, and primary retinal cells. Intravitreally administered fluorescent EVs were rapidly cleared from the vitreous. Retinal ganglion cells (RGCs) had maximal EV fluorescence at 14 days post administration, and microglia at 7 days. Both in vivo and in the explant model, most EVs were no deeper than the inner nuclear layer. Retinal astrocytes, microglia, and mixed neurons in vitro endocytosed EVs in a dose-dependent manner. Thus, our results indicate that intravitreal EVs are suited for the treatment of retinal diseases affecting the inner retina. Modification of the EV surface should be considered for maintaining EVs in the vitreous for prolonged delivery.

scholarly journals Transforming growth factor beta suppresses human immunodeficiency virus expression and replication in infected cells of the monocyte/macrophage lineage.

1991 ◽  
Vol 173 (3) ◽  
pp. 589-597 ◽  
Author(s):  
G Poli ◽  
A L Kinter ◽  
J S Justement ◽  
P Bressler ◽  
J H Kehrl ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
T Cell ◽  
Growth Factor ◽  
Cell Line ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Tgf Beta ◽  
Immunodeficiency Virus ◽  
Growth Factor Beta

The pleiotropic immunoregulatory cytokine transforming growth factor beta (TGF-beta) potently suppresses production of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome, in the chronically infected promonocytic cell line U1. TGF-beta significantly (50-90%) inhibited HIV reverse transcriptase production and synthesis of viral proteins in U1 cells stimulated with phorbol myristate acetate (PMA) or interleukin 6 (IL-6). Furthermore, TGF-beta suppressed PMA induction of HIV transcription in U1 cells. In contrast, TGF-beta did not significantly affect the expression of HIV induced by tumor necrosis factor alpha (TNF-alpha). These suppressive effects were not mediated via the induction of interferon alpha (IFN-alpha). TGF-beta also suppressed HIV replication in primary monocyte-derived macrophages infected in vitro, both in the absence of exogenous cytokines and in IL-6-stimulated cultures. In contrast, no significant effects of TGF-beta were observed in either a chronically infected T cell line (ACH-2) or in primary T cell blasts infected in vitro. Therefore, TGF-beta may play a potentially important role as a negative regulator of HIV expression in infected monocytes or tissue macrophages in infected individuals.

scholarly journals Differential Regulation of Mouse B Cell Development by Transforming Growth Factor β1

2002 ◽  
Vol 9 (2) ◽  
pp. 86-95 ◽  
Author(s):  
Denise A. Kaminski ◽  
John J. Letterio ◽  
Peter D. Burrows
Keyword(s):  
B Cells ◽  
Growth Factor ◽  
B Cell ◽  
Transforming Growth Factor ◽  
Plasma Cells ◽  
Population Analysis ◽  
Cell Development ◽  
B Cell Development

Transforming growth factor β (TGFβ) can inhibit thein vitroproliferation, survival and differentiation of B cell progenitors, mature B lymphocytes and plasma cells. Here we demonstrate unexpected, age-dependent reductions in the bone marrow (BM) B cell progenitors and immature B cells in TGFβ1-/-mice. To evaluate TGFβ responsiveness during normal B lineage development, cells were cultured in interleukin 7 (IL7)±TGFβ. Picomolar doses of TGFβ1 reduced pro-B cell recoveries at every timepoint. By contrast, the pre-B cells were initially reduced in number, but subsequently increased compared to IL7 alone, resulting in a 4-fold increase in the growth rate for the pre-B cell population. Analysis of purified BM sub-populations indicated that pro-B cells and the earliest BP1-pre-B cells were sensitive to the inhibitory effects of TGFβ1. However, the large BP1+pre-B cells, although initially reduced, were increased in number at days 5 and 7 of culture. These results indicate that TGFβ1 is important for normal B cell developmentin vivo, and that B cell progenitors are differentially affected by the cytokine according to their stage of differentiation.

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