Immortalization without neoplastic transformation of human mesenchymal stem cells by transduction with HPV16 <i>E6/E7</i> genes

Hung, Shih Chieh; Yang, Da; Chang, Che Shoa; Lin, Ruey‐Jen; Wang, Jih Shiuan; Ho, Larry L.T.; Yang, Wei‐Pang

doi:10.1002/ijc.20126

Cited by 87 publications

(92 citation statements)

References 37 publications

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“…To examine cell-cell interactions during chondrogenesis, immortalized human articular chondrocytes were obtained from the hPi cell line (4), and immortalized human MSCs (from bone marrow) were obtained from the Kp-hMSC line as previously described (35). Both cell lines were transduced with E6/E7 genes of human papillomavirus 16 and have been previously characterized for their proliferation and differentiation potentials.…”

Section: Cell Linesmentioning

confidence: 99%

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In vitro stage‐specific chondrogenesis of mesenchymal stem cells committed to chondrocytes

Chen

Lai

et al. 2009

Arthritis & Rheumatism

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110

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Objective. Osteoarthritis is characterized by an imbalance in cartilage homeostasis, which could potentially be corrected by mesenchymal stem cell (MSC)-based therapies. However, in vivo implantation of undifferentiated MSCs has led to unexpected results. This study was undertaken to establish a model for preconditioning of MSCs toward chondrogenesis as a more effective clinical tool for cartilage regeneration.Methods. A coculture preconditioning system was used to improve the chondrogenic potential of human MSCs and to study the detailed stages of chondrogenesis of MSCs, using a human MSC line, Kp-hMSC, in commitment cocultures with a human chondrocyte line, hPi (labeled with green fluorescent protein [GFP]). In addition, committed MSCs were seeded into a collagen scaffold and analyzed for their neocartilage-forming ability.Results. Coculture of hPi-GFP chondrocytes with Kp-hMSCs induced chondrogenesis, as indicated by the increased expression of chondrogenic genes and accumulation of chondrogenic matrix, but with no effect on osteogenic markers. The chondrogenic process of committed MSCs was initiated with highly activated chondrogenic adhesion molecules and stimulated cartilage developmental growth factors, including members of the transforming growth factor ␤ superfamily and their downstream regulators, the Smads, as well as endothelial growth factor, fibroblast growth factor, insulin-like growth factor, and vascular endothelial growth factor. Furthermore, committed Kp-hMSCs acquired neocartilageforming potential within the collagen scaffold. Osteoarthritis (OA), one of the most common musculoskeletal diseases, has been ascribed to an imbalance in cartilage homeostasis in the aging process (1,2). Hyaline cartilage has little capacity for self-repair. As a result, continuous mechanical stress can lead to the degradation of articular cartilage, culminating in a vicious cycle of destructive processes (3). Many therapeutic interventions directed at the restoration of the reparative capacity of chondrocytes have been explored (3-5). One of the emerging approaches is cell-based therapy, in which expanded chondrocytes are used for cartilage repair. However, the outcome of this approach has been disappointing, due to the difficulties in maintaining chondrocytic phenotypes and the decrease in proliferative capacity of autologous chondrocytes with increasing age (1,6,7). Conclusion. These findings help define the molecular markers of chondrogenesis and more accuratelyCell therapy based on autologous mesenchymal stem cells (MSCs), which have a vast proliferative ca-

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Section: Cell Linesmentioning

confidence: 99%

“…We have previously investigated the methods of purification, characterization, differentiation, and application of multipotent MSCs (10,(32)(33)(34)(35). We have also studied the expression of matrix proteins, the type I and type II collagens, which induce chondrogenesis in human MSCs under the influence of TGF␤1 (3).…”

mentioning

confidence: 99%

In vitro stage‐specific chondrogenesis of mesenchymal stem cells committed to chondrocytes

Chen

Lai

et al. 2009

Arthritis & Rheumatism

Self Cite

110

118

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“…Given that KP cells have been transfected using exogenous genes to maintain MSC stemness,28, 29 whether the incorporated genes produce different responses in these cells remain unclear. To test the inhibitory effects of Midazolam in chondrogenesis, we performed primary hMSCs to evaluate the results that were obtained from the KP MSC cell line.…”

Section: Resultsmentioning

confidence: 99%

“…KP cells, an immortalized human MSC line, were established from primary human MSCs, which were transduced using a retroviral vector LXSN‐16E6E7 28, 29. KP cells display similar types of cell surface marker profiles as parental primary human MSCs do and retain the stem‐like properties.…”

Section: Methodsmentioning

confidence: 99%

“…KP cells display similar types of cell surface marker profiles as parental primary human MSCs do and retain the stem‐like properties. KP cells can not only renew themselves, but also differentiate into mesenchymal and nonmesenchymal cell lineages without neoplastic transformation under certain conditions 28. This type of cell line has been used in studies of stem cell biology and regenerative medicine 30.…”

Section: Methodsmentioning

confidence: 99%

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Midazolam inhibits chondrogenesis via peripheral benzodiazepine receptor in human mesenchymal stem cells

Chen

Huang

et al. 2018

J Cellular Molecular Medi

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Midazolam, a benzodiazepine derivative, is widely used for sedation and surgery. However, previous studies have demonstrated that Midazolam is associated with increased risks of congenital malformations, such as dwarfism, when used during early pregnancy. Recent studies have also demonstrated that Midazolam suppresses osteogenesis of mesenchymal stem cells (MSCs). Given that hypertrophic chondrocytes can differentiate into osteoblast and osteocytes and contribute to endochondral bone formation, the effect of Midazolam on chondrogenesis remains unclear. In this study, we applied a human MSC line, the KP cell, to serve as an in vitro model to study the effect of Midazolam on chondrogenesis. We first successfully established an in vitro chondrogenic model in a micromass culture or a 2D high‐density culture performed with TGF‐β‐driven chondrogenic induction medium. Treatment of the Midazolam dose‐dependently inhibited chondrogenesis, examined using Alcian blue‐stained glycosaminoglycans and the expression of chondrogenic markers, such as SOX9 and type II collagen. Inhibition of Midazolam by peripheral benzodiazepine receptor (PBR) antagonist PK11195 or small interfering RNA rescued the inhibitory effects of Midazolam on chondrogenesis. In addition, Midazolam suppressed transforming growth factor‐β‐induced Smad3 phosphorylation, and this inhibitory effect could be rescued using PBR antagonist PK11195. This study provides a possible explanation for Midazolam‐induced congenital malformations of the musculoskeletal system through PBR.

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Fabrication of polycaprolactone scaffolds filled with cell‐compatible porous matrix for enhanced cell growth for biomedical applications

Hsu

Lin

et al. 2023

J of Applied Polymer Sci

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In this study, polycaprolactone (P) scaffolds are fabricated by 3D‐printing method, and subsequently a mixed solution of natural‐biocompatible chitosan (C), gelatin (G), and hydroxyapatite (H) is filled into the pores of P scaffolds. Freeze‐gelation method is then carried out to successfully form a cell‐compatible porous CGH matrix within P scaffold pores, obtaining a newly fabricated scaffold called CGH/P composite scaffold which has large specific surface area favorable for cell attachment and growth. The effects of slow cooling (sc) and fast cooling (fc) modes utilized in freeze process on pore structures of CGH matrix are observed via SEM. The use of fc mode creates a CGH matrix with a laminar structure which benefits nutrient permeation and cell migration. Mechanical properties tests indicate that the maximum compressive strength and Young's modulus of CGH/P‐fc scaffolds are 2–7 and 15–35 MPa, respectively. Cell compatibility tests show that on day 7, the cell numbers in the CGH/P‐fc scaffolds are about twofold of those in the P scaffolds, and the cell activities in the CGH/P‐fc scaffolds are even higher, indicating that the presence of laminar porous CGH matrix within P scaffold pores substantially enhances the migration, attachment, and growth of cells in the scaffolds. The results suggest that the fabricated CGH/P‐fc composite scaffold is highly promising for biomedical applications such as bone repair in the future.

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Immortalization without neoplastic transformation of human mesenchymal stem cells by transduction with HPV16 E6/E7 genes

Cited by 87 publications

References 37 publications

In vitro stage‐specific chondrogenesis of mesenchymal stem cells committed to chondrocytes

In vitro stage‐specific chondrogenesis of mesenchymal stem cells committed to chondrocytes

Midazolam inhibits chondrogenesis via peripheral benzodiazepine receptor in human mesenchymal stem cells

Fabrication of polycaprolactone scaffolds filled with cell‐compatible porous matrix for enhanced cell growth for biomedical applications

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