Evidence for histogenic interactions during in vitro limb chondrogenesis

Solursh, Michael; Reiter, Rebecca S.

doi:10.1016/0012-1606(80)90324-3

Cited by 61 publications

(26 citation statements)

References 21 publications

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“…Preliminary experiments revealed these conditions permit a linear uptake of radiolabel into acid-insoluble material. Solursh and Reiter [26] have shown a positive correlation between chondrogenesis and uptake of Na2 35SO4 in vitro, and DeLuca et al [29] have shown that 92% of the [35SO4 ~] incorporated by cultures of embryonic cartilage is into chondroitin sulfate.…”

Section: Methodsmentioning

confidence: 99%

Arachidonate metabolism during chondrogenesisin vitro

Chepenik

Waite

et al. 1984

Calcif Tissue Int

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Chick embryo limb bud mesenchyme cells undergoing chondrogenesis in vitro were labeled with [3H] arachidonic acid and [14C] palmitic acid, and stimulated by mechanical means to convert a portion of their incorporated [3H] to radiolabeled compounds which co-chromatographed with authentic prostaglandins in the appropriate thin layer chromatography system. Chondrogenesis was (1) inhibited by concentrations of indomethacin or eicosa-5,8,11,14-tetraynoic acid which inhibited conversion of [3H] to prostaglandinlike compounds; and (2) stimulated by prostaglandin E2. We interpret these data to mean that (1) cells undergoing chondrogenesis in vitro are able to metabolize endogenous arachidonic acid to prostaglandins, and (2) synthesis of prostaglandinlike compounds is requisite to chondrogenesis in vitro.

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Section: Methodsmentioning

confidence: 99%

Arachidonate metabolism during chondrogenesisin vitro

Chepenik

Waite

et al. 1984

Calcif Tissue Int

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“…cell condensation (21). Several factors have been shown to play a role in this process, including cell-cell interactions (22)(23)(24)(25)(26)(27), composition of the ECM (25, 28 -30), changes in cell shape (31), and response to cytokines (32). Following cell condensation, chondrogenic cells that produce type I collagen, fibronectin (FN), and its integrin receptor ␣ 5 ␤ 1 , differentiate to stage I chondrocytes that express type II collagen and eventually to stage II hypertrophic chondrocytes, characterized by type II and X collagen production (12).…”

mentioning

confidence: 99%

Integrins α6Aβ1 and α6Bβ1 Promote Different Stages of Chondrogenic Cell Differentiation

Segat¹,

Comai²,

Marco³

et al. 2002

Journal of Biological Chemistry

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The differentiation of chondrocytes and of several other cell types is associated with a switch from the ␣ 6B to the ␣ 6A isoform of the laminin ␣ 6 ␤ 1 integrin receptor. To define whether this event plays a functional role in cell differentiation, we used an in vitro model system that allows chick chondrogenic cells to remain undifferentiated when cultured in monolayer and to differentiate into chondrocytes when grown in suspension culture. We report that: (i) upon over-expression of the human ␣ 6B , adherent chondrogenic cells differentiate to stage I chondrocytes (i.e. increased type II collagen, reduced type I collagen, fibronectin, ␣ 5 ␤ 1 and growth rate, loss of fibroblast morphology); (ii) the expression of type II collagen requires the activation of p38 MAP kinase; (iii) the over-expression of ␣ 6A induces an incomplete differentiation to stage I chondrocytes, whereas no differentiation was observed in ␣ 5 and mock-transfected control cells; (iv) a prevalence of the ␣ 6A subunit is necessary to stabilize the differentiated phenotype when cells are transferred to suspension culture. Altogether, these results indicate a functional role for the ␣ 6B to ␣ 6A switch in chondrocyte differentiation; the former promotes chondrocyte differentiation, and the latter is necessary in stabilizing the differentiated phenotype.Growth factors, cell-extracellular matrix (ECM), 1 and cellcell interactions are the primary determinants of lineage decisions and differentiation events in embryogenesis. These regulatory events involve different types of receptors and may lead to activation of several signaling pathways, such as those mediated by MAP kinases (1-3). Changes in the expression pattern of most of these receptors, including the integrins (heterodimeric ␣␤ receptors involved in cell-ECM and cell-cell interactions), are able to modulate several events associated with cell differentiation (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20).The onset of chondrogenesis in developing long bones is characterized by the reduction of intercellular spaces and establishment of extensive cell-cell contacts between mesenchymal chondrogenic cells, i.e. cell condensation (21). Several factors have been shown to play a role in this process, including cell-cell interactions (22-27), composition of the ECM (25, 28 -30), changes in cell shape (31), and response to cytokines (32). Following cell condensation, chondrogenic cells that produce type I collagen, fibronectin (FN), and its integrin receptor ␣ 5 ␤ 1 , differentiate to stage I chondrocytes that express type II collagen and eventually to stage II hypertrophic chondrocytes, characterized by type II and X collagen production (12).Most of these events can be reproduced in vitro in a tissue culture model system that allows condensation and differentiation of chick embryo tibiae chondrogenic cells (12,(33)(34)(35). These cells, which adhere to tissue culture dishes and display a fibroblast-like phenotype (pre-chondrogenic cells), proliferate and secrete type I collagen...

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“…While other culture systems provide for expression of a chondrogenic phenotype (Solursh and Reiter, 1980;Archer et al, 1985;Kosher and Rodgers, 1987;Cottrill et al, 1987), our data suggest that the approach used in this study is unique in that it provides for the isolation of a subpopulation of mesoderm that not only retains its potential for overt chondrogenic cytodifferentiation, but also its potential for the formation of a predictable distribution ("pattern") of chondrogenic elements. Moreover, for the collective reasons discussed below, we have interpreted the in vitro formation of chondrogenic elements to suggest that the cells of the central subset retain reproducible characteristics of "limbness.…”

Section: Discussionmentioning

confidence: 78%

“…We define "limbness" according to Zwilling (1968) as not merely the potential of limb mesoderm to express a chondrogenic phenotype but by its predisposition to form a proximal to distal sequence of differentiated chondrogenic elements whose number changes with age in a manner characteristic of in situ limb mesoderm. Because homotypic and heterotypic cell interactions seem to be essential for chondrogenesis in the limb (Solursh and Reiter, 1980;Archer et al, 19851, we considered the maximal retention of naturally occurring cell-to-cell associations as a critical component of the culture approach. Indeed, as Frederick and Fallon (1982) have shown, recombinant limbs may lose their limb-like characteristics when dissociated anterior and posterior mesodermal cells are intermixed prior to grafting.…”

mentioning

confidence: 99%

Leg bud mesoderm retains morphogenetic potential to express limb‐like characteristics (“limbness”) in collagen gel culture

Isokawa

Krug

Fallon

et al. 1992

Developmental Dynamics

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Recent in situ hybridization studies have correlated expression of potential regulatory genes with pattern formation in limb bud mesoderm (Tabin: Cell 66:199-217,1991); however, the mechanism(s) controlling their expression in mesoderm and their relevance to the establishment of a limb morphogenetic pattern remain unknown. One likely candidate for regulating patterning events in limb mesoderm is the apical ectodermal ridge, as its removal in ovo results in a graded truncation of limb skeletal elements in the proximal-distal axis dependent upon the time of excision (Rowe and Fallon: J Embryo1 Exp Morph 68:l-7, 1982). In the present study, we investigate whether the hypothetical imprint of ridge ectoderm is retained in cultured mesoderm. Specifically, we sought to determine if a subpopulation of limb mesoderm that forms in collagen gel culture (Markwald et al: Anat Rec 226:91-107, 1990), retains any expression of "limbness" in the absence of limb ectoderm as characterized by the formation of a predictable number and distribution of limb-like chondrogenic elements in comparison to the temporal and spatial relationships of the in situ proximal, hindlimb skeletal structures. Accordingly, explants of undissociated mesoderm from stage 18-22 chicken leg buds were cultured without ectoderm on collagen gel lattices and the central subpopulation of mesoderm was examined morphologically. We show that this central subset of mesoderm will form chondrogenic cells which were not expressed uniformly throughout the subset, but rather distinct nodules or elements of cartilage were elaborated. Moreover, the number of elements expressed by the central subset increased with the age of the mesoderm at the time of explantation; spatially and temporally, the sequence of elements that formed always proceeded from the proximal, anterior margin of the subset to its distal, posterior border. The shapes of the initial elements (designated I and 11) resembled the forms of in situ proximal skeletal structures (girdle and femur-like), whereas more distal elements (111-V) were often fused and without structural similarity to in situ skeletal structures. frequency of element I formation was reduced approximately one-half, whereas formation of more distal elements was unaffected. Conversely, element formation from the central subset established from isolated anterior mesoderm was virtually identical to intact mesoblasts, indicating a capacity to regulate for the loss of mesoderm as occurs in situ (Hampe: Archs Anat Microsc MorDh Exp 48:345-378, 1959).We interpret these findings to mean that limb mesoderm which forms the central subset is not merely competent to express in the absence of continuous ectodermal co-culture, a chondrogenic phenotype, but also retains intrinsic, Zimblike morphogenetic potential to form, regulate, and, to a variable extent, shape a reproducibly specific number of cartilage elements. Because the number of chondrogenic elements that formed in culture appeared stage-specific, a role for the apical ridge in specifyin...

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Evidence for histogenic interactions during in vitro limb chondrogenesis

Cited by 61 publications

References 21 publications

Arachidonate metabolism during chondrogenesisin vitro

Arachidonate metabolism during chondrogenesisin vitro

Integrins α6Aβ1 and α6Bβ1 Promote Different Stages of Chondrogenic Cell Differentiation

Leg bud mesoderm retains morphogenetic potential to express limb‐like characteristics (“limbness”) in collagen gel culture

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