Ariane Rochat's research while affiliated with École Polytechnique Fédérale de Lausanne and other places
Publications (36)
Single-cell cloning is essential in stem cell biology, cancer research, and biotechnology. Regulatory agencies now require an indisputable proof of clonality that current technologies do not readily provide. Here, we report a one-step cloning method using an engineered pipet combined with an impedance-based sensing tip. This technology permits the...
Supplementary Table S1
There is a widespread agreement from patient and professional organisations alike that the safety of stem cell therapeutics is of paramount importance, particularly for ex vivo autologous gene therapy. Yet current technology makes it difficult to thoroughly evaluate the behaviour of genetically corrected stem cells before they are transplanted. To...
Review Process File
Supplementary Information
Replying to: T.-T. Sun, S. C. Tseng & R. M. Lavker 463, 10.1038/nature08805 (2010)Our claim is not that there are no stem cells in the limbus, but that there is more to corneal renewal than the limbus and that the double-dome-shaped structure of the cornea and physical constraints have a crucial impact on cell dynamics.
The integrity of the cornea, the most anterior part of the eye, is indispensable for vision. Forty-five million individuals worldwide are bilaterally blind and another 135 million have severely impaired vision in both eyes because of loss of corneal transparency; treatments range from local medications to corneal transplants, and more recently to s...
Stem cells safeguard tissue homeostasis and guarantee tissue repair throughout life. The decision between self-renewal and differentiation is influenced by a specialized microenvironment called stem cell niche. Physical and molecular interactions with niche cells and orientation of the cleavage plane during stem cell mitosis control the balance bet...
Skin protects the body against environmental hazards. It constantly renews and remodels, also repairing itself when wounded. Transplantation of autologous cultured epidermal stem cells has been part of the therapeutic arsenal to treat extensive burn wounds since the mid-eighties [1]. Permanently engrafted keratinocyte stem cells reconstitute the ep...
Given their accessibility, multipotent skin-derived cells might be useful for future cell replacement therapies. We describe the isolation of multipotent stem cell-like cells from the adult trunk skin of mice and humans that express the neural crest stem cell markers p75 and Sox10 and display extensive self-renewal capacity in sphere cultures. To d...
Given their accessibility, multipotent skin-derived cells might be useful for future cell replacement therapies. We describe the isolation of multipotent stem cell-like cells from the adult trunk skin of mice and humans that express the neural crest stem cell markers p75 and Sox10 and display extensive self-renewal capacity in sphere cultures. To d...
Adult stem cells are essential for tissue renewal, regeneration, and repair, and their expansion in culture is of paramount importance for regenerative medicine. Using the whisker follicle of the rat as a model system, we demonstrate that (i) clonogenicity is an intrinsic property of the adult stem cells of the hair follicle; (ii) after cultivation...
A challenge in ex vivo skin cell therapy is to obtain a massive expansion of the stem cells from a small skin biopsy in the shortest time possible, while preserving stemness. Cell therapy using autologous keratinocyte stem cells, cultivated according to the preceding method, has saved the lives of hundreds of burned patients, has restored sight to...
The upper region of the outer root sheath of vibrissal follicles of adult mice contains multipotent stem cells that respond to morphogenetic signals to generate multiple hair follicles, sebaceous glands, and epidermis, i.e., all the lineages of the hairy skin. At the time when hair production ceases and when the lower region of the follicle undergo...
We describe here eleven different mutations in SPINK5, encoding the serine protease inhibitor LEKTI, in 13 families with Netherton syndrome (NS, MIM256500). Most of these mutations predict premature termination codons. These results disclose a critical role of SPINK5 in epidermal barrier function and immunity, and suggest a new pathway for high ser...
Marie-Unna hypotrichosis (MU) is a rare autosomal dominant congenital alopecia characterised by progressive hair loss starting in early childhood, often aggravated at puberty and leading to scarring alopecia of variable severity. We have studied three multigeneration families of Belgian, British and French descent. The human genome was screened wit...
Epidermolysis bullosa simplex (EBS) arises from mutations within the keratin 5 and 14 (K5 and K14) genes which alter the integrity of basal keratinocytes cytoskeleton. The majority of these defects are missense mutations in the rod domain, whose locations influence the disease severity. We investigated a large family dominantly affected with the Do...
We describe two familial cases of dominant dystrophic epidermolysis bullosa (DDEB) that are heterozygous for deletions in COL7A1 that alter splicing, despite intact consensus splice-site sequences. One patient shows a 28-bp genomic deletion (6081del28) in exon 73 associated with the activation of a cryptic donor splice site within this exon; the co...
We have characterized 21 mutations in the type VII collagen gene (COL7A1) encoding the anchoring fibrils, 18 of which were not previously reported, in patients from 15 unrelated families with recessive dystrophic epidermolysis bullosa (RDEB). COL7A1 mutations in both alleles were identified by screening the 118 exons of COL7A1 and flanking intron r...
We have investigated 8 patients from 7 unrelated families with lamellar ichthyosis (LI) for defects in the keratinocyte transglutaminase (TGK) gene. We have characterized three novel homozygous mutations and a previously reported splice acceptor site mutation. One patient showed a C-to-T change in the binding site for the transcription factor Sp1 w...
Papillomaviruses are small DNA tumor viruses with a life cycle inseparably linked to the differentiation of the pluristratified epithelium. The infection of epithelial layers of the skin may remain latent or may result in the development of benign tumors. A certain number of distinct papillomavirus types, however, cause lesions which have a high ri...
Three subunits (alpha, beta, gamma) of the amiloride-sensitive epithelial sodium channel have been recently characterized. The channel subunits have significant homologies with the Caenorhabditis elegans mec-4, mec-10 and deg-1 genes, which are involved in control of cell volume and mecanotransduction. These subunits are coexpressed at equivalent l...
We have examined the growth capacity of keratinocytes isolated from human scalp hair follicles. Like the keratinocytes of glabrous epidermis, most of the colony-forming cells are classified as holoclones or meroclones when analyzed in a clonal assay. Some of them have extensive growth potential, as they are able to undergo at least 130 doublings. T...
The Hallopeau-Siemens type of recessive dystrophic epidermolysis bullosa (HS-RDEB) is a life-threatening autosomal disease characterized by loss of dermal-epidermal adherence with abnormal anchoring fibrils (AF). We recently linked HS-RDEB to the type VII collagen gene (COL7A1) which encodes the major component of AF. We describe a patient who is h...
The epidermis and its related appendages such as the hair follicle constitute the epithelial compartment of the skin. The exact location and distribution of the keratinocyte colony-forming cells within the epidermis or its appendages are unknown. We report that in the rat vibrissa, keratinocyte colony-forming cells are highly clustered in the bulge...
Epidermolysis bullosa simplex (EBS) is a group of epidermal blistering diseases almost invariably transmitted as a dominant trait, which has recently been shown to arise from mutations in keratins 14 and 5 (K14 and K5). We describe a family with recessive EBS in which the disease is tightly linked to the substitution of the highly conserved glutami...
Citations
... Tumor viruses have been suspected to target stem cells for transformation (61)(62)(63), and the Rb protein is commonly inhibited by tumor viruses (64). High-risk HPVs have been proposed to infect stem cells (65,66). A recent study showed that HPV16 preferentially transforms basal keratinocytes with stem cell properties (67) and that the HPV E7 oncoprotein, which inhibits Rb similarly to MCV LT, activates pluripotency genes, including Oct4, Sox2, and Nanog (68). ...
... The bulge area located at the level of insertion of the arrector pili muscle is considered to be a site of pluripotent epithelial stem cells [66]. This hypothesis was supported by the results of transplantation experiments in which isolated keratin 15-positive bulge stem cells were grafted to analyse cell fate plasticity, and it was demonstrated that they were capable of forming all cell linages, including hair follicles and sebaceous glands [19,66,68,72]. However, bulge stem cells are not the only cell source for sebaceous gland renewal. ...
... Diseases of the conjunctiva can therefore have significant ocular morbidity. Interestingly, there has been considerable debate regarding the role of the conjunctiva in corneal regeneration, and some have proposed that corneal and conjunctival epithelia are equipotent (Majo et al, 2010). Despite these findings and its essential role in ocular health, conjunctival stem cell biology has not been as well investigated as that of corneal stem cells (). ...
Reference: Stem Cells in Ophthalmology
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... As a result, clonal conversion is an irreversible phenomenon in normal condition [43] which leads to restriction on proliferative potential of KSCs [45]. Therefore, it's essential that gain an insight into the molecular mechanism that control clonal conversion in order to regenerate epidermis by development of cell- based therapeutic approaches for treatment of skin disorders [46] [42,47]. Furthermore, Vollmers et al. reported that isolated holoclones are able to create more percentage of holoclones in contrast to isolated meroclones [47]. ...
... Mutations in the TGM 1 gene resulting in reduced TGase 1 activity were identified soon after ( Russell et al. 1995). To date over 30 different mutations have been identified in the TGM 1 gene by several research groups (Study I and II) (Bichakjian et al. 1998; Huber et al. 1997; Parmentier et al. 1995; Petit et al. 1997; Pigg et al. 1998; Tok et al. 1999). Most of these are point mutations or small deletions resulting in amino acid substitutions, stop codons, or splicing defects. ...
Reference: Autosomal Recessive Congenital Ichthyosis
... In the future, the ability to expand significant numbers of human epithelial cells may also open the door to gene editing, mRNA, oligonucleotide, and/or cell therapy approaches in PCD. Notably, cell therapies using epithelial cells expanded on 3T3-J2 feeder layers are approved for clinical use for severe burns and limbal stem cell deficiency [34- 36] and expansion from single cell-derived clones is achievable [37]. However, as cellular therapy in genetic lung disease remains a distant prospect [38], model systems that enable the testing of gene therapies or small molecule approaches are much needed. ...
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... Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C-treated mouse 3T3-J2 fibroblasts. The co-culture was incubated at 37 °C with 10% CO 2 in a 3:1 mixture of Dulbecco-Vogt modification of Eagle's medium (DMEM) and Ham's F12 medium, supplemented with 10% fetal bovine serum (FBS), 1.8 × 10 −4 M adenine hemisulfate salt, 5 μg/ml insulin, 0.4 μg/ml hydrocortisone, 10 −10 M cholera toxin, and 2 × 10 −9 M triiodothyronine (T3), as described previously 1,30 . Keratinocytes between passages 2 and 7 were used for experiments, and the medium was changed every 4 days. ...
... When SCs are taken outside their native niche, they often undergo a stress response (16,62), a process analogous to wound repair in vivo (35,63). Of note, such stress is reversible in young HFSCs, and when transplanted back into mice, they resolve the stress and give rise to all lineages of HFs and epidermis (39,64). ...
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... L'EBD est due à une perte de l'adhésion entre l'épiderme et le derme sous-jacent liée à des mutations du gène COL7A1 (Hilal et al., 1993, Hovnanian et al., 1992 . Les patients présentent une prédisposition aux cancers cutanés (Fine et al., 2009). ...
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... Dowling-Meara 131760 AD (Coulombe et al., 1991;Hut et al., 2000;Pfendner et al., 2005a) Generalized intermediate EBS (formerly, Koebner type) 131900 AD (Bonifas, Rothman, & Epstein, 1991;Humphries et al., 1993;Pfendner et al., 2005b) Localized EBS (formerly, Weber-Cockayne) 131800 AD (Chen, Bonlfas, Matsumura, Blumenfeld & Epstein, 1993) Autosomal recessive EBS 601001 AR (Y. M. Hovnanian et al., 1993;Rugg et al., 1994) ITGB4 Integrin β4 EBS junctional is with pyloric atresia 226730 AR (Chavanas et al., 1999;Vidal et al., 1995) localized EBS (formerly, Weber-Cockayne) 131800 AD (Jonkman, Pas, Nijenhuis, Kloosterhuis, & Steege, 2002) PLEC1 Plectin EBS with muscular dystrophy 226670 AR (Bolling, Pas et al., 2010;Smith et al., 1996) EBS (Betz et al., 2006;Jones & Grice, 1978;Sprecher et al., 2007) EBS with migratory circinate erythema ...
Reference: Keratins and epidermolysis bullosa simplex
