The pulp organ is retained in the pulp chamber of teeth and maintains the biological and physiological vitality of the surrounding dentin. It works as a biosensor and generates secondary dentine and tertiary dentine to resist tooth abrasion and pathogenic stimuli (Zhang and Yelick, 2010). However, dental pulp is vulnerable to injury (Smulson and Sieraski, 1989). Most people experience some irreversible pulpal diseases during their lifetime. Hence, pulp regeneration is one of the research tasks in dentistry that attracts much attention. Stem cell transplantation is a plausible strategy for the regeneration of dental pulp organ. Dental pulp stem cells (DPSCs)derived from heavy or inflamed dental pulps have the natural advantage in pulp regeneration due to its dentinogenic potentiality (Huang et al., 2009). DPSCs are delivered into prepared root canal, which then differentiate into odontoblasts, fibroblasts, and other kinds of cells. It was shown that these transplanted DPSCs were able to produce dentin and formed a dentin-pulp like tissue both in vitro and in vivo(Huang, 2009).However, low survival rate of the transplanted cells is a common problem in pulp regeneration. Overexpression of Bcl-2 could enhance cell anti-apoptotic ability. Studies of many kinds of cell transplantation showed that a large number of cells died upon grafting and a large proportion of cell death seemed to have occurred due to apoptosis (Liu et al., 2013; Zhang et al., 2001).The aim of this study was to improve cell survival through making DPSCs overexpress lymphoma 2 (Bcl-2) protein.Bcl-2 is a proto-oncogene which playsa significant role in (anti) apoptosis. Former studies in the literature have provided evidences that overexpressing Bcl-2 could reduce cell apoptosis. However, this strategy has not been studied in the modification of DPSCs. In this study, DPSCs were isolated from discarded third molars of adults and manipulated to overexpressing Bcl-2. Proliferation of modified DPSCs was analyzed by static batch culture, CCK-8 test and BrdU based proliferation test. Apoptosis of modified DPSCs was analyzed by measuring DNA fragments in the cells. Modified DPSCs generated a higher maximum cell population during static batch culture and showed higher viability (the ratio of live cells to total cells). CCK-8 test showed that the population of modified DPSCs increased faster than control group cells and wild type cells. Modified DPSCs were not better than the other cells in proliferative ability, but had lower apoptosis level when culturing in serum free medium. Hence, overexpressing Bcl-2 could increase cell population, the mechanism is to help DPSCs survival rather than promote the proliferative ability of cells. / published_or_final_version / Dentistry / Master / Master of Philosophy

No description available.

No description available.

Τα τελευταία χρόνια υπάρχει μεγάλο ενδιαφέρον για τη βιολογία των σωματικών βλαστικών κυττάρων (adult stem cells) εξαιτίας της ικανότητας τους να αυτοανανεώνονται καθώς επίσης και της πλαστικότητας την οποία εμφανίζουν. Οι ιδιότητες αυτές καθιστούν τα σωματικά βλαστικά κύτταρα ικανά να παράγουν προγόνους διαφόρων τύπων ωρίμων κυττάρων, οι οποίοι συμμετέχουν ενεργά στη διατήρηση της ομοιόστασης ιστών και οργάνων τόσο κατά τη φυσιολογική πορεία της ζωής τους όσο και σε περίπτωση βλάβης. Εν γένει τα εμβρυικά και σωματικά βλαστικά κύτταρα φαίνεται να εμφανίζουν όμοιες λειτουργίες όσον αφορά στην ικανότητα αυτοανανέωσης και στην ιδιότητα τους να παράγουν διαφοροποιημένους κυτταρικούς προγόνους διαφόρων κυτταρικών σειρών τόσο σε απλοποιημένες συνθήκες καλλιέργειας in vitro όσο και μετά από μεταμόσχευσή τους σε ξενιστή in vivo. Πιο συγκεκριμένα η εδραίωση των ιδιοτήτων των βλαστικών και προγονικών κυττάρων in vitro και in vivo παρείχε τις ενδείξεις της συμμετοχής τους στην ανανέωση των ιστών μετά από βλάβη. Για το λόγο αυτό η χρήση τους αποτελεί πολλά υποσχόμενη στρατηγική στην κυτταρική και γενετική θεραπευτική πολλών εκφυλιστικών νοσημάτων καθώς επίσης και ως επικουρική ανοσοθεραπεία για διαφορους επιθετικούς τύπους καρκίνου. Η νόσος του Parkinson, η νόσος του Alzheimer, οι μυικές εκφυλιστικές διαταραχές, χρόνια ηπατικά νοσήματα, η καρδιακή ανεπάρκεια, ο σακχαρώδης διαβήτης τύπου I και τύπου II καθώς επίσης και διάφορες δερματικές, οφθαλικές, νεφρικές και αιμοποιητικές διαταραχές θα μπορούσαν να αντιμετωπιστούν με θεραπείες βασιζόμενες σε βλαστικά κύτταρα. Γενετικές μεταβολές ή/και επίμονη ενεργοποίηση διακριτών αναπτυξιακών μονοπατιών οι οποίες είναι δυνατόν να παρατηρούνται σε μειοψηφία σωματικών βλαστικών και προγονικών κυττάρων μπορούν, σε ορισμένες περιπτώσεις, να οδηγήσουν σε ογκογόνο μεταλλαγή. Το φαινόμενο αυτό εμπλέκει την ενεργοποίηση κυτταρικών μονοπατιών τα οποία σηματοδοτούνται από διάφορους αυξητικούς παράγοντες και ενέχουν σημαντική θέση στην αύξηση και επιβίωση καρκινικών κυττάρων. Πιο συγκεκριμένα η θετική ρύθμιση μονοπατιών που αφορούν στον EGF, hedgehog, Wnt/β-κατενίνη και του Νοtch αποτελούν κρίσιμης σημασίας γεγονότα για την έναρξη και προαγωγή επιθετικών τύπων κακοήθειας όπως η οξεία λευχαιμία και το λέμφωμα, ο καρκίνος του εγκεφάλου, του δέρματος, του πνεύμονα, του οισοφάγου, του στομάχου, του παγκρέατος, του ήπατος, του μαστού, των ωοθηκών, του προστάτη και των όρχεων. Για το λόγο αυτό η μοριακή στόχευση αυτών των ογκογόνων σηματοδοτικών στοιχείων αποτελεί πολλά υποσχόμενη προσέγγιση για την ανάπτυξη νέων θεραπευτικών συνδιασμών εναντίων μεταστατικών μορφών καρκίνου. / Stem cell biology and their role in lung homeostasis, tissue repair and cancer.

Skin-derived precursors (SKPs) are multipotent dermal stem cells that reside within a hair follicle niche and that share properties with embryonic neural crest precursors. Here, we have asked whether SKPs and their endogenous dermal precursors originate from the neural crest or whether, like the dermis itself, they originate from multiple developmental origins. To do this, we used two different mouse Cre lines that allow us to perform lineage tracing: Wnt1-cre, which targets cells deriving from the neural crest, and Myf5-cre, which targets cells of a somite origin. By crossing these Cre lines to reporter mice, we show that the endogenous follicle-associated dermal precursors in the face derive from the neural crest, and those in the dorsal trunk derive from the somites, as do the SKPs they generate. Despite these different developmental origins, SKPs from these two locations are functionally similar, even with regard to their ability to differentiate into Schwann cells, a cell type only thought to be generated from the neural crest. Analysis of global gene expression using microarrays confirmed that facial and dorsal SKPs exhibit a very high degree of similarity, and that they are also very similar to SKPs derived from ventral dermis, which has a lateral plate origin. However, these developmentally distinct SKPs also retain differential expression of a small number of genes that reflect their developmental origins. Thus, an adult neural crest-like dermal precursor can be generated from a non-neural crest origin, a finding with broad implications for the many neuroendocrine cells in the body.

Both cDNA tag-based and DNA chip hybridization assays have revealed widespread transcriptional activity across mammalian genomes, providing a rich source of novel protein-coding and non-coding transcripts. Annotation and functional evaluation of this undefined transcriptome space represents a major step towards the comprehensive definition of biomolecules regulating the properties of living cells, including embryonic stem cells (ESCs) and their derivatives. In this study I analysed 87 rare mRNA transcripts from human ESCs that mapped uniquely to the human genome, in regions lacking evidence for known genes or transcripts. In addition, the transcripts appeared enriched in the hESC transcriptome as enumerated by serial analysis of gene expression (SAGE). Full-length transcripts corresponding to twelve novel LongSAGE tags were recovered and evaluated with respect to gene structure, protein-coding potential, and gene regulatory features. In addition, transcript abundance was compared between RNA isolated from undifferentiated hESCs and differentiated cells. Analysis of full-length transcripts revealed that the novel ORFs did not exceed a size of 129 amino acids and no matches were observed to well characterized protein domains. Interesting protein level predictions included small disulfide-bonded proteins, known members of which are important in a variety of biological processes. Transcripts evaluated for differential expression by real-time RT-qPCR (Reverse Transcription followed by real-time quantitative Polymerase Chain Reaction) were found to be variably expressed (0.2- to 4.5-fold) in Day-2 or Day-4 retinoic acid-induced differentiation cultures compared to undifferentiated hESCs. Relative quantitation using a universal reference RNA (derived from pooled adult tissues) showed large differences in novel transcript levels (0.002- to 35-fold) compared to hESCs. Collectively, these results provide a detailed analysis of a set of novel hESC transcripts and their abundance in early and adult differentiated cell types, both of which may advance our understanding of the transcriptional events governing stem cell behavior.

No description available.

No description available.

No description available.

Physiological electric fields (EFs) have been measured in many developing and regenerating systems and are modulated spatially, temporally and in magnitude.  Many types of cells polarize and migrate in specific directions when exposed to a small applied EF similar in magnitude to those found endogenously.  In the present investigation, an endogenous EF was recorded in an aortic explant.  The response of three EPCs (MFLM-4, AEL-deltaR1 and AEL-deltaR1/Runx1), one endothelial cell line (HUVEC) and five other cell lines (MDA-MB-231, MTLn-3, HEK-293, COS-7 and CHO) in physiological EFs were shown.  The roles of vascular endothelial growth factor receptor 2 (VEGFR2) and voltage-gated potassium (Kv) channels in the EFs directed cell migration were studied.  The roles of intracellular calcium, intracellular calcium stores, purinergic signalling and gap junction communication in the intracellular calcium rising, calcium wave propagation and re-endothelialization were studied too.  In conclusion, electrical signalling between cells and ionic fluxes in cells are important controlling mechanisms of endothelial behaviours.  Electric fields and ionic fluxes regulate migration of endothelial cells and endothelial progenitor cells.  Orchestrated endothelial behaviours and recruitment of endothelial progenitor cells repair injuries to endothelium, and may lead to angiogenesis.  Manipulation of electric fields and control of ionic fluxes may be a promising approach to control endothelial cell migration and angiogenesis.