nave-adult-stem-cells-isolation-from-primary-human-fibroblast-cultures

Na&iuml;ve Adult Stem Cells Isolation from Primary Human Fibroblast Cultures

Over the last decade, several adult stem cell  populations have been identified in human skin 1-4.  The isolation of multipotent adult dermal precursors ...

Technische Universität München

Netherton syndrome (NS, MIM 256500) is a potential live threatening autosomal-recessive skin disorder clinically characterized by the trias of congenital erythroderma, hair shaft anomalies and atopic diathesis. It is caused by mutations in the gene SPINK5 resulting in a deficiency of its processed protein named lympho-epithelial Kazal-type related inhibitor (LEKTI). LEKTI controls the activity of several serine proteases in the skin that are involved in terminal differentiation. Loss of LEKTI results in protease hyperactivity, increased degradation of intercellular junctions, reduced stratum corneum adhesion and impaired skin barrier function. Today NS can only be treated symptomatically.

rAAV2-mediated restoration of LEKTI in LEKTI-deficient cells from Netherton patients.

Syndrome de Netherton (NS, MIM 256500) est un d&eacute;sordre de peau autosomique r&eacute;cessive mena&ccedil;ant vivants potentiels cliniquement caract&eacute;ris&eacute; par le trias d&#39;Erythrodermie cong&eacute;nitale, anomalies arbre cheveux et diath&egrave;se atopique. Elle est caus&eacute;e par des mutations dans le g&egrave;ne SPINK5, ce qui entra&icirc;ne une d&eacute;ficience de ses prot&eacute;ines transform&eacute;es nomm&eacute; Kazal-type lympho-&eacute;pith&eacute;lial associ&eacute;es inhibiteur (LEKT1). LEKT1 contr&ocirc;le l&#39;activit&eacute; de plusieurs prot&eacute;ases de s&eacute;rine dans la peau qui sont impliqu&eacute;s dans la diff&eacute;renciation terminale. Perte de LEKT1 r&eacute;sultats dans l&#39;hyperactivit&eacute; de la prot&eacute;ase, a augment&eacute; de d&eacute;gradation des jonctions intercellulaires, r&eacute;duit l&#39;adh&eacute;rence de la couche corn&eacute;e et alt&eacute;ration de la fonction barri&egrave;re de la peau. Aujourd&#39;hui NS ne peuvent &ecirc;tre trait&eacute;es selon les sympt&ocirc;mes.

restauration induite par le rAAV2 de LEKT1 dans les cellules d&eacute;ficientes en LEKT1 de Netherton patients.

Sindrome di Netherton (NS, MIM 256500) &egrave; una malattia autosomica recessiva pelle live potenziale minaccioso caratterizzata clinicamente da trias di eritrodermia congenita, anomalie dell&#39;albero dei capelli e diatesi atopica. &Egrave; causata da mutazioni nel gene SPINK5 con conseguente deficit delle sue proteine trasformate denominato lympho-epitelio Kazal-tipo relativi inibitore (LEKTI). LEKTI controlla l&#39;attivit&agrave; di diverse proteasi della serina della pelle che sono coinvolti nella differenziazione terminale. Perdita di LEKTI risultati iperattivit&agrave; proteasi, aumentata degradazione delle giunzioni intercellulari, ridotto strato corneo adesione e alterata funzione barriera della pelle. Oggi NS possono essere trattate solo sintomaticamente.

restauro rAAV2-mediato di LEKTI nelle cellule carenti LEKTI da Netherton pazienti.

Hutchinson-Gilford progeria syndrome (HGPS, OMIM 176670) is a rare disorder characterized by segmental accelerated aging and early death from coronary artery disease or stroke. Nearly 90% of HGPS sufferers carry a G608G mutation within exon 11 of LMNA, producing a truncated form of prelamin A, referred to as "progerin". Here, we report the isolation of naÃ¯ve multipotent skin-derived precursor (SKP) cells from dermal fibroblast cultures from HGPS donors. These cells form spheres and express the neural crest marker, nestin, in addition to the multipotent markers, OCT4, Sox2, Nanog and TG30; these cells can self-renew and differentiate into smooth muscle cells (SMCs) and fibroblasts. The SMCs derived from the HGPS-SKPs accumulate nuclear progerin with increasing passages. A subset of the HGPS-naÃ¯ve SKPs express progerin in vitro and in situ in HGPS skin sections. This is the first in vivo evidence that progerin is produced in adult stem cells, and implies that this protein could induce stem cells exhaustion as a mechanism contributing to aging. Our study provides a basis on which to explore therapeutic applications for HGPS stem cells and opens avenues for investigating the pathogenesis of other genetic diseases.

NaÃ¯ve adult stem cells from patients with Hutchinson-Gilford progeria syndrome express low levels of progerin in vivo.

Syndrome de progeria de Hutchinson-Gilford (hormones de croissance, OMIM 176670) est une affection rare caract&eacute;ris&eacute;e par vieillissement acc&eacute;l&eacute;r&eacute; segmentaire et la mort pr&eacute;coce de la maladie coronarienne ou d&#39;accident vasculaire c&eacute;r&eacute;bral. Pr&egrave;s de 90 % des personnes souffrant d&#39;hormones de croissance portent une mutation du g&egrave;ne LMNA, produisant une forme tronqu&eacute;e de l&#39;identification du prelamin A, d&eacute;nomm&eacute; &laquo; progerin &raquo; G608G exon 11. Nous rapportons ici, l&#39;isolement des cellules multipotentes d&eacute;riv&eacute;es de peau pr&eacute;curseur (SKP) na&iuml;ve des cultures de fibroblastes dermiques de donateurs d&#39;hormones de croissance. Ces cellules forment des sph&egrave;res et expriment le marqueur de la cr&ecirc;te neurale, nestin, outre les marqueurs multipotentes, OCT4, Sox2, Nanog et TG30 ; ces cellules peuvent auto-renouveler et se diff&eacute;rencient en cellules musculaires lisses (CML) et de fibroblastes. Le CML d&eacute;riv&eacute; les hormones de croissance-SKP accumule progerin nucl&eacute;aire avec une augmentation de passages. Un sous-ensemble des SKP d&#39;hormones de croissance-na&iuml;ve express progerin in vitro et in situ dans les sections de peau hormones de croissance. Il s&#39;agit de la premi&egrave;re preuve in vivo que progerin est produite dans les cellules souches adultes et implique que cette prot&eacute;ine pouvait induire &eacute;puisement de cellules souches comme un m&eacute;canisme qui contribuent au vieillissement. Notre &eacute;tude fournit une base explorer les applications th&eacute;rapeutiques pour les hormones de croissance des cellules souches et ouvre des pistes pour l&#39;&eacute;tude de la pathogen&egrave;se des autres maladies g&eacute;n&eacute;tiques.

Na&iuml;ve des cellules souches adultes provenant de patients atteints du syndrome de Hutchinson-Gilford progeria expriment des niveaux faibles de progerin in vivo.

Sindrome di progeria di Hutchinson-Gilford (HGPS, OMIM 176670) &egrave; una malattia rara caratterizzata da invecchiamento accelerato segmentale e morte precoce di malattia coronarica o ictus. Quasi il 90% dei malati di HGPS trasportano una mutazione nell&#39;esone 11 G608G LMNA, producendo una forma troncata di prelamin A, indicato come &quot;progerin&quot;. Riportiamo l&#39;isolamento delle cellule multipotenti derivate da pelle precursore (SKP) ingenuo da colture di fibroblasti dermici da donatori HGPS. Queste cellule formano sfere ed esprimono il marcatore cresta neurale, la nestina, oltre i marcatori multipotenti, OCT4, Sox2, Nanog e TG30; Queste cellule possono auto-rinnovarsi e differenziarsi in cellule muscolari lisce (SMC) e fibroblasti. SMCs derivato da HGPS-SKPs si accumulano progerin nucleare con l&#39;aumento di passaggi. Un sottoinsieme dei SKPs HGPS-na&iuml;ve esprimono progerin in vitro ed in situ in HGPS pelle sezioni. Questa &egrave; la prima prova in vivo che progerin &egrave; prodotto in cellule staminali adulte e implica che questa proteina potrebbe indurre cellule staminali esaurimento come un meccanismo che contribuiscono all&#39;invecchiamento. Il nostro studio fornisce una base esplorare le applicazioni terapeutiche per le cellule staminali HGPS e apre le vie per indagare la patogenesi di altre malattie genetiche.

Article	Year
rAAV2-mediated restoration of LEKTI in LEKTI-deficient cells from Netherton patients. Journal of dermatological science\| PubMed ID: 21251800	2011
NaÃ¯ve adult stem cells from patients with Hutchinson-Gilford progeria syndrome express low levels of progerin in vivo. Biology open\| PubMed ID: 23213444	2012

Daniela Roedl

Biography

JoVE Journal Publications

Other Publications