Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes

Ichinose, Shizuko; Muneta, Takeshi; Koga, Hideyuki; Segawa, Yuko; Tagami, Motoki; Tsuji, Kunikazu; Sekiya, Ichiro

doi:10.1038/labinvest.2009.125

Cited by 39 publications

(27 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Before sectioning, Epon block samples were prepared by cutting the enamel area. Ultrathin (90 nm) sections were collected on copper grids, double-stained with uranyl acetate and lead citrate, and then examined by TEM (H-7100, Hitachi, Tokyo, Japan) (Ichinose et al, 2010). Semi-thin (1 μm) sections were collected on glass slides and stained with Toluidine Blue.…”

Section: Transmission Electron Microscopy (Tem) and Toluidine Blue Stmentioning

confidence: 99%

Mohawk transcription factor regulates homeostasis of the periodontal ligament

et al. 2016

Self Cite

View full text Add to dashboard Cite

The periodontal ligament (PDL), which connects the teeth to the alveolar bone, is essential for periodontal tissue homeostasis. Although the significance of the PDL is recognized, molecular mechanisms underlying PDL function are not well known. We report that mohawk homeobox (Mkx), a tendon-specific transcription factor, regulates PDL homeostasis by preventing its degeneration. Mkx is expressed in the mouse PDL at the age of 10 weeks and expression remained at similar levels at 12 months. In Mkx −/− mice, agedependent expansion of the PDL at the maxillary first molar (M1) furcation area was observed. Transmission electron microscopy (TEM) revealed that Mkx −/− mice presented collagen fibril degeneration in PDL with age, while the collagen fibril diameter gradually increased in Mkx +/+ mice. PDL cells lost their shape in Mkx −/− mice, suggesting changes in PDL properties. Microarray and quantitative polymerase chain reaction (qPCR) analyses of Mkx −/− PDL revealed an increase in osteogenic gene expression and no change in PDL-and inflammatory-related gene expression. Additionally, COL1A1 and COL1A2 were upregulated in Mkxoverexpressing human PDL fibroblasts, whereas osteogenic genes were downregulated. Our results indicate that Mkx prevents PDL degeneration by regulating osteogenesis.

show abstract

Section: Transmission Electron Microscopy (Tem) and Toluidine Blue Stmentioning

confidence: 99%

Mohawk transcription factor regulates homeostasis of the periodontal ligament

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Stemness indicators, among them the morphological features, are essential in order to differentiate stem cells capable of addressing transplantation procedures from defective cell lines unusable for further applications (Pascucci et al, 2010). Up to now, the fine morphology of MSCs has been described in only a few reports regarding the bone marrow of different species, amnion and chorionic term placenta and human adult blood vessels (Csaki et al, 2007;Ichinose et al, 2010;Karaoz et al, 2009;Pascucci et al, 2010;Pasquinelli et al, 2007).…”

Section: Microscopy Research and Techniquementioning

confidence: 99%

Ultrastructural analysis of human bone marrow mesenchymal stem cells during in vitro osteogenesis and chondrogenesis

Teti¹,

Cavallo²,

Grigolo³

et al. 2011

Microscopy Res & Technique

View full text Add to dashboard Cite

The main purpose of this article was to describe the morphology of mesenchymal stem cells (MSCs) differentiated in vitro towards osteogenic and chondrogenic lineages and to focus on the ultrastructural features associated with these processes. Human mononuclear cells (hMNC) were isolated, expanded, and analyzed for the expression of specific cell surface markers to demonstrate their stem cell characteristics. Human mononuclear cells were differentiated in vitro in an osteogenic and in a chondrogenic sense for 7, 14, 21, and 28 days. Subsequently, they were processed using electron microscopic analysis (FEISEM). Alizarin red and alcian blue staining were carried out to demonstrate the deposition of mineral salts and proteoglycans in the extracellular matrix. Undifferentiated MSCs showed a cell surface covered by filopodia and ondulopodia. During differentiation, the MSCs changed their shape from a round to a fibroblastic-like shape. At the end of the differentiation, several filaments with a parallel orientation in the osteogenic samples as well as a network organization in the chondrogenic samples were detected in the extracellular spaces. This study demonstrated that there are morphological features associated with the undifferentiated and differentiated states of the MSCs, which could be utilized as new parameters for identifying and classifying these cells.

show abstract

“…Although cell nuclei were scarce, pellets from both, control and chondrogenic medium had abundant ECM and specifi c structures (islets) that resembled hyaline cartilage made of collagen type II. No comparable cartilage-like tissue has been described in literature although several papers described high chondrogenic potential of SF cells comparing to adipose tissue and bone marrow [11,29]. Interestingly, synovial fl uid progenitors expressing CD90+ from normal but not osteoarthritic joints undergo chondrogenic differentiation without micro-mass culture [17].…”

Section: Discussionmentioning

confidence: 99%

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

et al. 2015

View full text Add to dashboard Cite

The aim of this study was to determine whether synovial fl uid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37ºC, 5% CO 2 , humidifi ed atmosphere) three weeks before their phenotype was characterized by fl ow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets' slides have shown a signifi cant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.

show abstract

Morphological differences during in vitro chondrogenesis of bone marrow-, synovium-MSCs, and chondrocytes

Cited by 39 publications

References 25 publications

Mohawk transcription factor regulates homeostasis of the periodontal ligament

Mohawk transcription factor regulates homeostasis of the periodontal ligament

Ultrastructural analysis of human bone marrow mesenchymal stem cells during in vitro osteogenesis and chondrogenesis

Proliferation And Differentiation Potential Of Canine Synovial Fluid Cells

Contact Info

Product

Resources

About