Regulation of proliferation and migration in retinoic acid treated C3H10T1/2 cells by TGF‐β isoforms

Makhijani, Nalini S.; Bischoff, David S.; Yamaguchi, Dean T.

doi:10.1002/jcp.20128

Cited by 23 publications

(19 citation statements)

References 59 publications

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“…When we analyzed the transcription of single receptor types, we found that RA treatment of either CLP-SP fibroblasts or N-SP cells downregulated TGFBR1, upregulated TGFBR2, and did not affect TGFBR3 gene expression. Although a previous study (49) reported RA decreases mRNA expression of TGF-β receptors, the significance of our differing results with each receptor is impossible to establish within the scope of the present study. In the final analysis, RA treatment of both CLP-SP and N-SP fibroblasts appears to modulate TGF-β3 system activity, which could account for the altered ECM composition (less GAG and FN production) that we found in RA-treated cells.…”

Section: Discussioncontrasting

confidence: 72%

Retinoic Acid, GABA-ergic, and TGF-β Signaling Systems Are Involved in Human Cleft Palate Fibroblast Phenotype

Baroni

Bellucci

Lilli

et al. 2006

Mol Med

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During embryogenesis, a complex interplay between extracellular matrix (ECM) molecules, regulatory molecules, and growth factors mediates morphogenetic processes involved in palatogenesis. Transforming growth factor-β (TGF-β), retinoic acid (RA), and γ-aminobutyric acid (GABA)ergic signaling systems are also potentially involved. Using ]methionine incorporation, anion exchange chromatography, semiquantitative radioactive RT-PCR, and a TGF-β binding assay, we aimed to verify the presence of phenotypic differences between primary cultures of secondary palate (SP) fibroblasts from 2-year-old subjects with familial nonsyndromic cleft lip and/or palate (CLP-SP fibroblasts) and age-matched normal SP (N-SP) fibroblasts. The effects of RA-which, at pharmacologic doses, induces cleft palate in newborns of many species-were also studied. We found an altered ECM production in CLP-SP fibroblasts that synthesized and secreted more glycosaminoglycans (GAGs) and fibronectin (FN) compared with N-SP cells. In CLP-SP cells, TGF-β3 mRNA expression and TGF-β receptor number were higher and RA receptor-α (RARA) gene expression was increased. Moreover, we demonstrated for the first time that GABA receptor (GABRB3) mRNA expression was upregulated in human CLP-SP fibroblasts. In N-SP and CLP-SP fibroblasts, RA decreased GAG and FN secretion and increased TGF-β3 mRNA expression but reduced the number of TGF-β receptors. TGF-β receptor type I mRNA expression was decreased, TGF-β receptor type II was increased, and TGF-β receptor type III was not affected. RA treatment increased RARA gene expression in both cell populations but upregulated GABRB3 mRNA expression only in N-SP cells. These results show that CLP-SP fibroblasts compared with N-SP fibroblasts exhibit an abnormal phenotype in vitro and respond differently to RA treatment, and suggest that altered crosstalk between RA, GABAergic, and TGF-β signaling systems could be involved in human cleft palate fibroblast phenotype.

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Section: Discussioncontrasting

confidence: 72%

Retinoic Acid, GABA-ergic, and TGF-β Signaling Systems Are Involved in Human Cleft Palate Fibroblast Phenotype

Baroni

Bellucci

Lilli

et al. 2006

Mol Med

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“…Thus, the functional motility characteristics we found for cells in our study agree with the fluorescence-activated cell sorting (FACS) profiling of the human cells indicating that these bone marrow-derived MSCs display an uncommitted phenotype, as is the case for marrow-derived MSCs from human and rabbit in the absence of exogenous osteogenic factors (51,66). Furthermore, our results suggest that at least rabbit and human cells have a similar motogenic behavior, and do not display the interspecies cytokine-induced motogenic variability that has been seen when comparing mice, rat, and human MSCs for TGF-␤ (16,43,44,65), an inconsistency that has also been reflected with other nonosteogenic fibroblastic cells (48).…”

Section: Fig 4 Dose-response For Haptotaxis Of Rabbit and Humansupporting

confidence: 52%

“…The parallel use of rabbit and human cells was judged necessary on the basis of previous literature suggesting interspecies extrapolation to be misleading. The in vitro variability of phenotype markers such as alkaline phosphatase production and osteoinduction potential comparing MSCs from different species can be significant (42), as can chemotaxis comparing mice/rat (17,43,44) and human (16) MSCs, using conserved factors such as transforming growth factor-␤ (TGF␤1) and fibroblast growth factor-2 (FGF-2).…”

Section: Introduction Mmentioning

confidence: 99%

Fibronectin, Vitronectin, and Collagen I Induce Chemotaxis and Haptotaxis of Human and Rabbit Mesenchymal Stem Cells in a Standardized Transmembrane Assay

Thibault

Hoemann

Buschmann

2007

Stem Cells and Development

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The mesenchymal stem cell (MSC) is a critical element in tissue repair and regeneration. Its ability to differentiate into multiple connective tissue cell types and to self-renew has made it a prime candidate in regenerative medicine strategies. Currently, the environmental cues responsible for in situ recruitment and control of MSC distribution at repair sites are not entirely revealed and in particular the role of extracellular matrix (ECM) proteins as motogenic factors has not been studied. Here we have used a standardized transmembrane chemotaxis assay to assess the chemotactic and haptotactic potential of fibronectin, vitronectin, and collagen type 1 on MSCs from both rabbit and human origin. The use of both cell types was based in part on the widespread use of rabbit models for musculoskeletal-related tissue engineering and repair models and their unknown correspondence to human in terms of MSC migration. The optimized assay yielded a greatly increased chemotactic response toward known factors such as platelet-derived growth factor-BB (PDGF)-BB compared to previous studies. Our primary finding was that all three ECM proteins tested (fibronectin, vitronectin, and collagen I) induced significant motogenic activity, in both soluble and insoluble forms, for both rabbit and human MSCs. These results suggest that ECM proteins could play roles as significant as cytokines in the recruitment of pluripotential repair cells wound and tissue repair sites. Furthermore, designed ECM coatings of scaffolds or implants could provide a new tool to control both cell influx and outflux from the scaffold post-implantation. Finally, the similarity of motogenic behavior of both rabbit and human cells suggests the rabbit is a reliable model for assessing MSC recruitment in repair and regeneration strategies. 489

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“…20 Cells (2.5×10 4 in 100 μL) were plated in triplicate onto Costar transwell membranes in the serum-free HMSCGM with 0.1% bovine serum albumin. Lower chambers contained HMSCGM with 1% fetal bovine serum (FBS).…”

Section: Methodsmentioning

confidence: 99%

Constitutive Expression of Human Telomerase Enhances the Proliferation Potential of Human Mesenchymal Stem Cells

Bischoff

Makhijani

Yamaguchi

2012

BioResearch Open Access

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Human mesenchymal stem cells (hMSCs) are highly desirable cells for bone engineering due to the inherent multipotent nature of the cells. Unfortunately, there is a high degree of variability, as primary hMSC cultures quickly undergo replicative senescence with loss of proliferative potential as they are continually propagated in cell culture. We sought to reduce the variability of these cells by insertion and expression of human telomerase reverse transcriptase (TERT) to immortalize the cell line. hMSCs were transduced with a lentivirus containing the human TERT gene. The resulting cell line has been propagated through more than 70 population-doubling level (PDL) to date and continues to grow exhibiting the characteristic fibroblastic hMSC phenotype. Expression of TERT mRNA and protein activity was confirmed in the TERT-transduced cells. Mock-transduced hMSCs had almost undetectable levels of TERT mRNA and protein activity and lost proliferation potential at PDL 14. The enhanced growth capacity of the hMSC TERT cells was due to increased cell proliferation and reduced cellular senescence rather than due to inhibition of apoptosis. The multipotent nature of the TERT cells was confirmed by differentiation toward the osteoblastic and adipogenic lineages in vitro. Osteoblastic differentiation was confirmed by both expression of alkaline phosphate and mineral deposition visualized by Alizarin Red staining. Adipogenic differentiation was confirmed by production of lipid droplets, which were detected by Oil Red-O staining. In summary, we have generated a stable hMSC line that can be continually propagated and retains both osteoblastic and adipogenic differentiation potential.

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Regulation of proliferation and migration in retinoic acid treated C3H10T1/2 cells by TGF‐β isoforms

Cited by 23 publications

References 59 publications

Retinoic Acid, GABA-ergic, and TGF-β Signaling Systems Are Involved in Human Cleft Palate Fibroblast Phenotype

Retinoic Acid, GABA-ergic, and TGF-β Signaling Systems Are Involved in Human Cleft Palate Fibroblast Phenotype

Fibronectin, Vitronectin, and Collagen I Induce Chemotaxis and Haptotaxis of Human and Rabbit Mesenchymal Stem Cells in a Standardized Transmembrane Assay

Constitutive Expression of Human Telomerase Enhances the Proliferation Potential of Human Mesenchymal Stem Cells

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