Neuropilin 1mediates keratinocyte growth factor signaling in adipose-derived stem cells: potential involvement in adipogenesis

Adipogenesis is regulated by a complex network of molecules, including fibroblast growth factors. Keratinocyte growth factor (KGF) has been previously reported to promote proliferation on rat preadipocytes, although the expression of its specific receptor, FGFR2-IIIb/KGFR, is not actually detected in mesenchymal cells. Here, we demonstrate that human adipose-derived stem cells (ASCs) show increased expression of KGF during adipogenic differentiation, especially in the early steps. Moreover, KGF is able to induce transient activation of ERK and p38 MAPK pathways in these cells. Furthermore, KGF promotes ASC differentiation and supports the activation of differentiation pathways, such as those of PI3K/Akt and the retinoblastoma protein (Rb). Notably, we observed only a low amount of FGFR2-IIIb in ASCs, which seems not to be responsible for KGF activity. Here, we demonstrate for the first time that Neuropilin 1 (NRP1), a transmembrane glycoprotein expressed in ASCs acting as a coreceptor for some growth factors, is responsible for KGF-dependent pathway activation in these cells. Our study contributes to clarify the molecular bases of human adipogenesis, demonstrating a role of KGF in the early steps of this process, and points out a role of NRP1 as a previously unknown mediator of KGF action in ASCs

ANCA-Negative Paucimmune Glomerulonephritis and Glomerular Recovery: Possible Role of Mesenchymal Stem Cells in a 69 Year-Old Patient

BACKGROUND: Pauci-immune crescentic glomerulonephritis is one of the most common causes of rapidly progressive glomerulonephritis, usually associated with anti-neutrophil cytoplasmic antibody (ANCA) but a minority of patients lack ANCAs. To date no effective treatments have been addressed for ANCA-negative glomerulonephritis despite several studies describe the possibility to adopt pharmacologic therapies. Cellular, molecular and preclinical studies demonstrate that mesenchymal stem cells (MSCs) contribute to glomerular cell turnover and repair and might represent a new therapeutic approach for ANCA-negative glomerulonephritis. AIM: In our study we presented a case report of a patient with ANCA-negative pauci-immune glomerulonephritis in order to underlie the growing need of an alternative therapy for glomerular diseases. MATERIALS AND METHODS: We report the case of a 69 year-old woman who presented to our department with diarrhea, vomiting and increased serum creatinine levels. At the First Aid Department, renal ultrasonography excluded dilatations of urinary tract and since the occurrence of progressive dyspnea, anuria and severe hypertension renal replacement therapy was started. RESULTS: A second kidney Doppler ultrasonography showed hyperechoic parenchyma, reduced thickness and increased resistive index. Urinalysis revealed persistent active sediment and proteinuria. Autoimmunity profile showed C3, IgE and IgM increased levels. Renal biopsy was performed showing diffuse extracapillary proliferative glomerulonephritis with negative Immunofluorescence (IF). ANCA test was negative for a second time by both IF analysis and antigen-specific ELISA. The diagnosis was ANCA-negative glomerulonephritis and steroid bolus was administered, improving urinary sediment and proteinuria but not glomerular filtration rate. DISCUSSION: Our data demonstrated that pharmacological treatment of ANCA-negative glomerulonephritis correlated with poor response and is not effective, pointing out the importance of new therapeutic options for this disease. In this regard MCSs may represent a valid alternative in the treatment of ANCA-negative glomerulonephritis. CONCLUSIONS: MSCs may provide an effective therapeutic option in glomerular diseases and future studies should be performed in order to pave the way for this cell-based therapy

Adipose-Derived Stem Cell: an Innovative Therapeutic Approach in Systemic Sclerosis and Parry-Romberg Syndrome

INTRODUCTION: Cell-based therapies represent a promising therapeutic approach to enhance the regeneration of damaged tissue and the combination with specific soluble mediators and biomaterial scaffolds has allowed the introduction of new treatment strategies in regenerative medicine. Adipose-derived stem cells (ASCs) show the ability to differentiate into several cell lineages such as chondrocytes, osteoblasts, adipocytes, neuronal cells, and muscle cells. These cells are abundant in normal human fat and they can be easily harvested from small amount of liposuction. For these reasons, they are greatly employed in the treatment of cutaneous and musculoskeletal defects are greatly. MATERIALS AND METHODS: ASCs isolated from liposuction aspirate by type I collagenases digestion were successively characterized by FACS analysis in order to verify their mesenchymal origin and test their capacity to differentiate into adipogenic, chondrogenic and osteogenic lineages. Autologous cult

Characterization of Human Vaginal Mucosa Cells for Autologous In Vitro Cultured Vaginal Tissue Transplantation in Patients with MRKH Syndrome

Mayer-Rokitansky-Küster-Hauser (MRKH) is a rare syndrome characterized by congenital aplasia of the uterus and vagina. The most common procedure used for surgical reconstruction of the neovagina is the McIndoe vaginoplasty, which consists in creation of a vaginal canal covered with a full-thickness skin graft. Here we characterized the autologous in vitro cultured vaginal tissue proposed as alternative material in our developed modified McIndoe vaginoplasty in order to underlie its importance in autologous total vaginal replacement. To this aim human vaginal mucosa cells (HVMs) were isolated from vaginal mucosa of patients affected by MRKH syndrome and characterized with respect to growth kinetics, morphology, PAS staining, and expression of specific epithelial markers by immunofluorescence, Western blot, and qRT-PCR analyses. The presence of specific epithelial markers along with the morphology and the presence of mucified cells demonstrated the epithelial nature of HMVs, important for an efficient epithelialization of the neovagina walls and for creating a functional vaginal cavity. Moreover, these cells presented characteristics of effective proliferation as demonstrated by growth kinetics assay. Therefore, the autologous in vitro cultured vaginal tissue might represent a highly promising and valid material for McIndoe vaginoplasty

Characterization of Human Vaginal Mucosa Cells for Autologous In Vitro Cultured Vaginal Tissue Transplantation in Patients with MRKH Syndrome

Mayer-Rokitansky-Küster-Hauser (MRKH) is a rare syndrome characterized by congenital aplasia of the uterus and vagina. The most common procedure used for surgical reconstruction of the neovagina is the McIndoe vaginoplasty, which consists in creation of a vaginal canal covered with a full-thickness skin graft. Here we characterized the autologous in vitro cultured vaginal tissue proposed as alternative material in our developed modified McIndoe vaginoplasty in order to underlie its importance in autologous total vaginal replacement. To this aim human vaginal mucosa cells (HVMs) were isolated from vaginal mucosa of patients affected by MRKH syndrome and characterized with respect to growth kinetics, morphology, PAS staining, and expression of specific epithelial markers by immunofluorescence, Western blot, and qRT-PCR analyses. The presence of specific epithelial markers along with the morphology and the presence of mucified cells demonstrated the epithelial nature of HMVs, important for an efficient epithelialization of the neovagina walls and for creating a functional vaginal cavity. Moreover, these cells presented characteristics of effective proliferation as demonstrated by growth kinetics assay. Therefore, the autologous in vitro cultured vaginal tissue might represent a highly promising and valid material for McIndoe vaginoplasty

Gene expression profile of patients with mayer- Rokitansky-küster- hauser syndrome: New insights into the potential role of developmental pathways

Mayer-Rokitansky-Küster-Hauser syndrome (MRKHS) is a rare disease characterized by congenital aplasia of uterus and vagina. Although many studies have investigated several candidate genes, up to now none of them seem to be responsible for the aetiology of the syndrome. In our study, we identified differences in gene expression profile of in vitro cultured vaginal tissue of MRHKS patients using whole-genome microarray analysis. A group of eight out of sixteen MRKHS patients that underwent reconstruction of neovagina with an autologous in vitro cultured vaginal tissue were subjected to microarray analysis and compared with five healthy controls. Results obtained by array were confirmed by qRT-PCR and further extended to other eight MRKHS patients. Gene profiling of MRKHS patients delineated 275 differentially expressed genes, of which 133 downregulated and 142 upregulated. We selected six deregulated genes (MUC1, HOXC8, HOXB2, HOXB5, JAG1 and DLL1) on the basis of their fold change, their differential expression in most patients and their relevant role in embryological development. All patients showed upregulation of MUC1, while HOXB2 and HOXB5 were downregulated, as well as Notch ligands JAG1 and DLL1 in the majority of them. Interestingly, HOXC8 was significantly upregulated in 47% of patients, with a differential expression only in MRKHS type I patients. Taken together, our results highlighted the dysregulation of developmental genes, thus suggesting a potential alteration of networks involved in the formation of the female reproductive tract and providing a useful clue for understanding the pathophysiology of MRKHS

List of 46 genes showing altered expression in both type I and type II MRKHS.

<p>List of 46 genes showing altered expression in both type I and type II MRKHS.</p

Summary of MRKHS patients.

<p>Summary of MRKHS patients.</p

Genes selected for qRT-PCR validation based on altered expression in microarray assay.

<p>Genes selected for qRT-PCR validation based on altered expression in microarray assay.</p