In vitro paracrine regulation of human keratinocyte growth by fibroblast‐derived insulin‐like growth factors

Barreca, Antonina; Luca, Michele De; Monte, P. Del; Bondanza, Sergio; Damonte, G.; Cariola, G; Marco, Eddi Di; Giordano, Giulio; Cancedda, Ranieri; Minuto, Francesco

doi:10.1002/jcp.1041510207

Cited by 142 publications

(85 citation statements)

References 23 publications

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“…23 The healing effects reported for IGF-1 have been attributed to a mitogenic effect on fibroblasts and keratinocytes, antiapoptotic effects via phosphatidylinositol-3 kinase signaling pathway, 24,25 and stimulation of local collagen formation. [26][27][28] In this study, liposomal IGF-1 cDNA gene transfer also prevents burn-induced increases in proinflammatory cytokine mRNA expression levels. The burn-induced elevated IL-1␤ mRNA levels present in the burn wound were attenuated by IGF-1-cDNA treatment.…”

Section: Discussionmentioning

confidence: 57%

“…22,23 The healing effects of IGF-1 have been attributed to mitogenic effects on fibroblasts and keratinocytes, anti-apoptotic effects via phosphatidylinositol-3 kinase signaling pathway, 24,25 and the stimulation of local collagen formation. [26][27][28] We tested the hypothesis that localized paracrine IGF-1 production, resulting from transient localized gene transfer of CMVdriven IGF-1-cDNA, ameliorates the prolonged local inflammation typically triggered by burn trauma and facilitates wound healing. In this study, local liposomal IGF-1 gene transfer is shown to attenuate increases in IL-1␤ and TNF-␣ and increase the levels of the anti-inflammatory cytokine IL-4 in the burn wound.…”

Section: Introductionmentioning

confidence: 99%

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Liposomal IGF-1 gene transfer modulates pro- and anti-inflammatory cytokine mRNA expression in the burn wound

et al. 2001

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Section: Discussionmentioning

confidence: 57%

Section: Introductionmentioning

confidence: 99%

Liposomal IGF-1 gene transfer modulates pro- and anti-inflammatory cytokine mRNA expression in the burn wound

et al. 2001

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“…There are numerous studies that have focused on the development of nutritionally optimized, readily defined, reproducible media and culture conditions for keratinocyte cells (26)(27)(28)(29). The aims of the present study were not to establish a new medium, and therefore, the Rheinwald and Green (30) protocol was applied.…”

Section: Discussionmentioning

confidence: 99%

Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin

et al. 2015

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Abstract. Currently, culture and growth keratinocytes are important stages in achieving a reliable and reproducible skin tissue. In the present study, two different methods, enzymatic and explant methods, for keratinocytes isolation from human foreskin were compared. Foreskins were cut into 2-3 mm pieces and placed in trypsin at 4˚C overnight for separation of the epidermis from the dermis. Subsequently, these samples were divided into two groups: i) Keratinocytes separated from the epidermis by trypsin and ii) by the explant method. These keratinocytes were divided into two groups: i) With no feeder layer and ii) onto a type I collagen scaffold. The cells were evaluated using immunocytochemistry and 4',6-diamidine-2'-phenylindole dihydrochloride (DAPI) staining. In the enzymatic treatment, after 7-10 days no attached cells were found in the cell culture dishes. In the explant method, keratinocytes were separated after ~24 h, attached rapidly and formed big colonies into a collagen scaffold. In the absence of a feeder layer, small colonies were developed with rapid loss of proliferation within 2-3 days. Keratinocytes showed positive immunoreactivity for the pan-cytokeratin marker and keratinocytes' nuclei were clearly observed. This method could be applied and developed as a component of skin substitutes to treat burns and wounds and also in laboratory testing. IntroductionThe skin is the largest organ in the body that is divided into two anatomically distinct regions, the dermis and epidermis. The normal structure and function of this organ is dependent on the intact epidermis anchored to its vascular, elastic dermis (1,2). Fibroblasts are the most prevalent cell types in the dermis, which produce different growth factors that induce proliferation of keratinocytes in vivo and in vitro (3). The principal cell type of the epidermis is the keratinocyte (1,4), which is a small epithelial cell, located at the top of the epidermal basal membrane and characterized by a low division rate (5,6).Thus far, various enzymatic methods for dermal-epidermal separation have been applied (6-8). For instance, trypsin separates suprabasal hemidesmosomes that causes the basal layer cells to attach to the dermal layer (6). Thermolysin is another enzyme that selectively separates desmosomes (5,9) and is able to separate the epidermis at the basal membrane zone level (5,10).Although the dissociation method of keratinocytes in primary culture is well-established, attempts to acquire purified adult stem-cell like⁄progenitor keratinocytes from whole human skin are still ongoing. In particular, different techniques are currently being applied to achieve high purity or homogeneous primary cultures enriched in keratinocyte progenitor⁄precursor cells. These include filtration, density gradient centrifugation and fluorescence-activated cell sorting using cell surface antibodies, as well as differential adhesion to enrich the cells that rapidly attach to particular substrates (11).A previous study showed that when keratinocytes/precursor cells...

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“…Although IGF-I increases keratinocyte activity in vitro (2,3,28) and overexpression or targeted disruption of the genes encoding IGF-I or IGF type 1 receptor (29,30) potently affect the dermis, little is still known about mechanisms that control production of IGF-I in this tissue.…”

Section: Discussionmentioning

confidence: 99%

“…In skin, IGF-I induces keratinocyte replication and skin matrix protein synthesis (2). However, keratinocytes do not express detectable amounts of IGF-I and may rely on the circulation, or perhaps expression by other local skin cells, for a supply of this factor (3,4). In contrast, keratinocytes express another growth regulator, parathyroid hormone-related protein (PTHrp), which shares amino-terminal sequence homology with PTH and acts at least in part through conventional PTH receptors (5,6).…”

mentioning

confidence: 99%

Parathyroid Hormone-related Protein Enhances Insulin-like Growth Factor-I Expression by Fetal Rat Dermal Fibroblasts

Shin

Casinghino

et al. 1997

Journal of Biological Chemistry

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Interactions between cells of differing embryonic origins comprise a common theme during tissue development and repair. Often, communication between them can be mediated by soluble growth mediators and in some cases is restricted in focus. That is, some cells respond to, but do not produce, mediators expressed by other cells within the tissue. Because keratinocytes respond to but do not express insulin-like growth factor I (IGF-I), another skin cell population, the dermal fibroblast, may supply this factor. However, keratinocytes express, but do not respond to parathyroid hormone related protein (PTHrp), which increases cAMP production by dermal fibroblasts. Based on earlier results where inducers of cAMP increase local IGF-I expression in skeletal tissue, we postulated that PTHrp might induce local IGF-I by dermal fibroblasts and provide a source of this factor for keratinocyte activity. Our studies reveal that IGF-I mRNA and protein levels increase in response to PTHrp in vitro, and that this effect is replicated by inducers of cAMP, but not by activators of protein kinase C. Consequently, these factors appear to comprise a paracrine loop within the skin, permitting focused but restricted IGF-I expression to support skin growth, remodeling, or repair.Insulin-like growth factor-I (IGF-I) 1 regulates a variety of actions in many somatic tissues (1). In skin, IGF-I induces keratinocyte replication and skin matrix protein synthesis (2). However, keratinocytes do not express detectable amounts of IGF-I and may rely on the circulation, or perhaps expression by other local skin cells, for a supply of this factor (3, 4). In contrast, keratinocytes express another growth regulator, parathyroid hormone-related protein (PTHrp), which shares amino-terminal sequence homology with PTH and acts at least in part through conventional PTH receptors (5, 6). PTHrp was first purified from squamous cell tumors where it was implicated in the paraneoplastic syndrome, humoral hypercalcemia of malignancy. It is a highly conserved and ubiquitous protein with many effects in a broad range of tissues (reviewed in Refs. 6 and 7).Our earlier studies first demonstrated that IGF-I expression is enhanced by PTH, PTHrp, prostaglandin E 2 , and other inducers of cyclic adenosine monophosphate (cAMP) in osteoblasts (8 -10). Despite the assumption that skin, and dermis in particular, is not a primary target organ for PTH, receptors shared by PTH and PTHrp are clearly present on dermal fibroblasts (11-13) in which fibronectin synthesis increases after exposure to PTHrp (14). Therefore, we postulated that keratinocyte-derived PTHrp might act as a local regulator in skin in a manner similar to that by circulating PTH in the skeleton. In this study we asked specifically if PTHrp could increase IGF-I production by fetal dermal fibroblasts through a cAMP-dependent pathway. In this way, keratinocytes might support dermal fibroblast function through the action of PTHrp, and as a result dermal fibroblasts could support keratinocyte activity through an incr...

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In vitro paracrine regulation of human keratinocyte growth by fibroblast‐derived insulin‐like growth factors

Cited by 142 publications

References 23 publications

Liposomal IGF-1 gene transfer modulates pro- and anti-inflammatory cytokine mRNA expression in the burn wound

Liposomal IGF-1 gene transfer modulates pro- and anti-inflammatory cytokine mRNA expression in the burn wound

Comparison of the enzymatic and explant methods for the culture of keratinocytes isolated from human foreskin

Parathyroid Hormone-related Protein Enhances Insulin-like Growth Factor-I Expression by Fetal Rat Dermal Fibroblasts

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