Chondrogenic differentiation of human pluripotent stem cells in chondrocyte co-culture

Qu, Chengjuan; Puttonen, Katja A.; Lindeberg, Heli; Ruponen, Marika; Hovatta, Outi; Koistinaho, Jari; Lammi, Mikko J.

doi:10.1016/j.biocel.2013.05.029

Cited by 64 publications

(50 citation statements)

References 66 publications

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“…The medium was changed three times per week during the culture period for all differentiation experiments. At the end of the differentiation periods, the cell pellets from chondrogenic differentiation were examined with histological stainings—for proteoglycans (PGs), with toluidine blue staining; for type II collagen, with immunohistochemistry [33,34]. The differentiated cells from osteogenesis and adipogenesis assays were visualized with stainings for alkaline phosphate (ALP) activity and Oil Red O (ORO), respectively [33,34].…”

Section: Methodsmentioning

confidence: 99%

Behavior of Human Bone Marrow-Derived Mesenchymal Stem Cells on Various Titanium-Based Coatings

Kaitainen

Kröger

et al. 2016

Materials

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The chemical composition and texture of titanium coatings can influence the growth characteristics of the adhered cells. An enhanced proliferation of the human mesenchymal stem cells (hMSCs) would be beneficial. The present study was aimed to investigate whether titanium deposited at different atmospheres would affect the cell growth properties, cellular morphology, and expression of surface markers of hMSCs. Titanium-based coatings were deposited on silicon wafers under oxygen, nitrogen, or argon atmospheres by ultra-short pulsed laser deposition using two different gas pressures followed by heating at 400 °C for 2 h. The characteristics of the coated surfaces were determined via contact angle, zeta potential, and scanning electron microscopy (SEM) techniques. Human MSCs were cultivated on differently coated silicon wafers for 48 h. Subsequently, the cell proliferation rates were analyzed with an MTT assay. The phenotype of hMSCs was checked via immunocytochemical stainings of MSC-associated markers CD73, CD90, and CD105, and the adhesion, spreading, and morphology of hMSCs on coated materials via SEM. The cell proliferation rates of the hMSCs were similar on all coated silicon wafers. The hMSCs retained the MSC phenotype by expressing MSC-associated markers and fibroblast-like morphology with cellular projections. Furthermore, no significant differences could be found in the size of the cells when cultured on all various coated surfaces. In conclusion, despite certain differences in the contact angles and the zeta potentials of various titanium-based coatings, no single coating markedly improved the growth characteristics of hMSCs.

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Section: Methodsmentioning

confidence: 99%

Behavior of Human Bone Marrow-Derived Mesenchymal Stem Cells on Various Titanium-Based Coatings

Kaitainen

Kröger

et al. 2016

Materials

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“…The finding that somatic cells can be reprogrammed back to the embryonic stem cell like induced pluripotent stem cells by the induced expressions of the certain combination of the four genes Oct3/4, Sox2, c‐Myc, and Klf4 or Oct4, Sox2, c‐Myc, and Nanog have attracted a lot of attention . However, this technique has not yet been therapeutically practical due to an insufficient knowledge of how to guide the differentiation to the desired cell types without inducing the formation of teratomas.…”

Section: Introductionmentioning

confidence: 99%

Rho‐kinase inhibitor Y‐27632 increases cellular proliferation and migration in human foreskin fibroblast cells

Piltti

Varjosalo

et al. 2015

Proteomics

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The idea of direct differentiation of somatic cells into other differentiated cell types has attracted a great interest recently. Rho-kinase inhibitor Y-27632 (ROCKi) is a potential drug molecule, which has been reported to support the gene expressions typical for the chondrocytes, thus restricting their phenotypic conversion to fibroblastic cells upon the cellular expansion. In this study, we have investigated the short-term biological responses of ROCKi to human primary foreskin fibroblasts. The fibroblast cells were exposed to 1 and 10 μM ROCKi treatments. A proteomics analysis revealed expression changes of 56 proteins, and a further protein pathway analysis suggested their association with the cell morphology, the organization, and the increased cellular movement and the proliferation. These functional responses were confirmed by a Cell-IQ time-lapse imaging analysis. Rho-kinase inhibitor treatment increased the cellular proliferation up to twofold during the first 12 h, and a wound model based migration assay showed 50% faster filling of the mechanically generated wound area. Additionally, significantly less vinculin-associated focal adhesions were present in the ROCKi-treated cells. Despite the marked changes in the cell behavior, ROCKi was not able to induce the expression of the chondrocyte-specific genes, such as procollagen α1 (II) and aggrecan.

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“…4 ng of cDNA was used in each qRT-PCR reaction. The specific primer sequences are presented in Supplementary Table 1 53–63 . Primers were purchased from Oligomer (Oligomer Oy) and from Invitrogen (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

Rho-kinase inhibitor Y-27632 and hypoxia synergistically enhance chondrocytic phenotype and modify S100 protein profiles in human chondrosarcoma cells

Piltti

Bygdell

Fernández-Echevarría

et al. 2017

Sci Rep

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Articular chondrocytes are slowly dividing cells that tend to lose their cell type-specific phenotype and ability to produce structurally and functionally correct cartilage tissue when cultured. Thus, culture conditions, which enhance the maintenance of chondrocyte phenotype would be very useful for cartilage research. Here we show that Rho-kinase inhibition by Y-27632 under hypoxic conditions efficiently maintains and even enhances chondrocyte-specific extracellular matrix production by chondrocytic cells. The effects of long-term Y-27632 exposure to human chondrosarcoma 2/8 cell phenotype maintenance and extracellular matrix production were studied at normoxia and at a 5% low oxygen atmosphere. Y-27632 treatment at normoxia induced ACAN and COL2A1 gene up-regulation and a minor increase of sulfated glycosaminoglycans (sGAGs), while type II collagen expression was not significantly up-regulated. A further increase in expression of ACAN and COL2A1 was achieved with Y-27632 treatment and hypoxia. The production of sGAGs increased by 65.8%, and ELISA analysis revealed a 6-fold up-regulation of type II collagen. Y-27632 also induced the up-regulation of S100-A1 and S100-B proteins and modified the expression of several other S100 protein family members, such as S100-A4, S100-A6, S100-A13 and S100-A16. The up-regulation of S100-A1 and S100-B proteins is suggested to enhance the chondrocytic phenotype of these cells.

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Chondrogenic differentiation of human pluripotent stem cells in chondrocyte co-culture

Cited by 64 publications

References 66 publications

Behavior of Human Bone Marrow-Derived Mesenchymal Stem Cells on Various Titanium-Based Coatings

Behavior of Human Bone Marrow-Derived Mesenchymal Stem Cells on Various Titanium-Based Coatings

Rho‐kinase inhibitor Y‐27632 increases cellular proliferation and migration in human foreskin fibroblast cells

Rho-kinase inhibitor Y-27632 and hypoxia synergistically enhance chondrocytic phenotype and modify S100 protein profiles in human chondrosarcoma cells

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