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- Fossum, Magdalena, et al.
(author)
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Long-term culture of human urothelial cells : a qualitative analysis
- 2005
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In: Cells Tissues Organs. - : S. Karger AG. - 1422-6405 .- 1422-6421. ; 181:1, s. 11-22
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Journal article (peer-reviewed)abstract
- Today, in vitro culturing of autologous cells is an established method in the field of tissue reconstruction. It can be applied to urothelial cells and could have many clinical implications in urological reconstructive surgery. This development calls for quality controls concerning cells used for clinical treatment when cells are autotransplanted back to the patient. We have studied cultured cells in order to detect whether genetic or morphologic changes occur. Urothelial cells isolated from bladder lavage were cultured according to different protocols based on the presence or absence of feeder cells. Genetic studies were performed by means of karyotyping with standard G-banding and interphase fluorescent in situ hybridization (FISH) analyses. The morphology of these epithelial cells was judged as well as immunostaining for epithelial cell markers. In addition, to minimize the risk of feeder cell contamination, proliferation studies were performed on cultures including feeder cells that had been pretreated with different doses of mitomycin or radiation. In initial studies, when using feeder cells in each passage according to standard protocols, urothelial cells proliferated unfavourably after the fourth passage with increasing numbers of mouse cells as well as urothelial tetraploid cells. We could also show that urothelial cells from bladder lavage need feeder cells in order to establish primary cultures. Further propagation up to 14 passages was performed without feeder cells and the urothelial cells retained normal karyotypes. We also found that mitomycin treatment had its main effect on feeder cells during the first 2 h. When feeder cells were irradiated, 20 Gy was effective and no feeder cell contamination was seen. In conclusion, we found that a high standard of quality in urothelial cell culturing can be achieved with a careful culturing technique.
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- Gustafson, Carl-Johan, et al.
(author)
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Employing human keratinocytes cultured on macroporous gelatin spheres to treat full thickness-wounds : an in vivo study on athymic rats.
- 2007
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In: Burns. - : Elsevier BV. - 0305-4179 .- 1879-1409. ; 33:6, s. 726-35
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Journal article (peer-reviewed)abstract
- Providing cutaneous wounds with sufficient epidermis to prevent infections and fluid loss is one of the most challenging tasks associated with surgical treatment of burns. Recently, application of cultured keratinocytes in this context has allowed this challenge to be met without several of the limitations connected with the use of split-thickness skin grafts. The continuous development of this novel approach has now revealed that transplantation of cultured autologous keratinocytes as single-cell suspensions exhibits several advantages over the use of cultured epidermal grafts. However, a number of methodological problems remain to be solved, primarily with regards to the complexity of culturing these cells; loss of viability and other negative effects during their preparation and transportation; the relatively long period of time required following transplantation to obtain a sufficiently protective epidermis. In the present investigation we attempted to eliminate these limitations by culturing the keratinocytes on macroporous gelatin spheres. Accordingly, the efficacies of normal human keratinocytes in single-cell suspension or growing on macroporous gelatin spheres, as well as of split-thickness skin grafts in healing wounds on athymic rats were compared. Human keratinocytes were found to adhere and proliferate efficiently both on the surface and within the pores of such spheres. Transplantation of such cells adherent to the spheres resulted in significantly more rapid formation of a stratified epidermis than did transplantation of single-cell suspensions or spheres alone. Twenty-three days after transplantation, the epidermis formed from the cells bound to the spheres was not as thick as the epidermis on wounds covered with split-thickness skin grafts, but significantly thicker than on wounds to which single-cell suspensions, spheres alone or no transplant at all was applied. Furthermore, fluorescence in situ hybridisation revealed that the transplanted keratinocytes, both those adherent to gelatin spheres and those in single-cell suspension, were components of the newly formed epidermis. These findings indicate that application of biodegradable macroporous spheres may prove to be of considerable value in designing cell-based therapies for the treatment of acute and persistent wounds.
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- Huss, Fredrik, 1971-, et al.
(author)
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Characterisation of a new degradable polymer scaffold for regeneration of the dermis : In vitro and in vivo human studies
- 2008
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In: Organogenesis. - 1547-6278. ; 4:3, s. 195-200
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Journal article (peer-reviewed)abstract
- Full thickness skin wounds in humans heal with scars, but without regeneration of the dermis. A degradable poly(urethane urea) scaffold (PUUR), Artelon® is already used to reinforce soft tissues in orthopaedics, and for treatment of osteoarthritis of the hand, wrist and foot. In this paper we have done in vitro experiments followed by in vivo studies to find out whether the PUUR is biocompatible and usable as a template for dermal regeneration. Human dermal fibroblasts were cultured on discs of PUUR, with different macrostructures (fibrous and porous). They adhered to and migrated into the scaffolds, and produced collagen. The porous scaffold was judged more suitable for clinical applications and 4 mm Ø, 2 mm-thick discs of porous scaffold (12% w/w or 9% w/w polymer solution) were inserted intradermally in four healthy human volunteers. The implants were well tolerated and increasing ingrowth of fibroblasts was seen over time in all subjects. The fibroblasts stained immunohistochemically for procollagen and von Willebrand factor, indicating neocollagenesis and angiogenesis within the scaffolds. The PUUR scaffold may be a suitable material to use as a template for dermal regeneration. ©2008 Landes Bioscience.
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- Junker, Johan, et al.
(author)
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Differentiation of human dermal fibroblasts towards endothelial cells
- 2013
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In: Differentiation. - Oxon, United Kingdom : Elsevier. - 0301-4681 .- 1432-0436. ; 85:3, s. 67-77
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Journal article (peer-reviewed)abstract
- The ultimate goal of vascular tissue engineering is the production of functional grafts for clinical use. Difficulties acquiring autologous endothelial cells have motivated the search for alternative cell sources. Differentiation of dermal fibroblasts towards several mesenchymal lineages as well as endothelial cells has been proposed. The aim of the present study was to investigate the endothelial differentiation capacity of human dermal fibroblasts on a gene expression, protein expression and functional physiological level. Endothelial differentiation of fibroblasts was induced by culturing cells in 30% human serum, but not in fetal calf serum. Expression of proteins and genes relevant for endothelial function and differentiation was increased after induction. Furthermore, fibroblasts exposed to 30% human serum displayed increased uptake of low-density lipoprotein and formation of capillary-like networks. The results of this study may have an impact on cell sourcing for vascular tissue engineering, and the development of methods for vascularization of autologous tissue engineered constructs.
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- Kjölhede, Anders, et al.
(author)
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Metoidioplasty and groin flap phalloplasty as two surgical methods for the creation of a neophallus in female-to-male gender-confirming surgery : A retrospective study comprising 123 operated patients.
- 2019
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In: JPRAS Open. - : Elsevier BV. - 2352-5878. ; 22, s. 1-8
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Journal article (peer-reviewed)abstract
- Background: In gender-confirming surgery of the female-to-male gender dysphoric patient, there is currently no ideal method for the creation of a neophallus. Historically, in our clinic, groin flap phalloplasty (GFP) has been the dominating method, but during the last 20 years, it has gradually been replaced with metoidioplasty (MP). The aim of this study was to investigate whether this change of method has influenced factors such as the frequency of complications and the number of operations needed to complete the reconstruction of the neophallus.Methods: This is a retrospective, single-centre, study comprising 123 consecutive female-to-male patients receiving a neophallus by GFP or MP between 2002 and 2015 at Linköping University Hospital, Sweden.Results: One-hundred twenty-three patients underwent 126 primary surgical procedures (39 GFPs and 87 MPs) with the intention of reconstructing a neophallus. The mean number of procedures required in the GFP group was 5.2 ± 2.7 compared with that of 2.4 ± 1.7 in the MP group (p < 0.001). In the GFP group, 18/39 (46.2%) had a documented complication compared with 30/87 (34.5%) in the MP group; however, the difference was not statistically significant (p = 0.21).Conclusions: The present study shows that the shift in method from GFP to MP has resulted in a decreased number of complications as well as a decrease in total surgical occasions. Both methods were found to be associated with relatively high frequencies of complications, however, mostly minor.
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