Kati Juuti-Uusitalo

Tampere University | UTA · Faculty of Medicine and Health Technology

The small intestinal epithelium has an important role in nutrition, but also in drug absorption and metabolism. There are a few two-dimensional (2D) patient-derived induced pluripotent stem cell (iPSC)-based intestinal models enabling easy evaluation of transcellular transport. It is known that animal-derived components induce variation in the expe...

In vitro cell culture models representing the physiological and pathological features of the outer retina are urgently needed. Artificial tissue replacements for patients suffering from degenerative retinal diseases are similarly in great demand. Here, we developed a co-culture system based solely on the use of human induced pluripotent stem cell (...

Diabetic retinopathy is a common diabetic complication that threatens the eye sight and may eventually lead to acquired visual impairment or blindness. While a substantial heritability has been reported for proliferative diabetic retinopathy (PDR), only a few genetic risk factors have been identified. Using genome-wide sib-pair linkage analysis inc...

In diabetic patients, high blood glucose induces alterations in retinal function and can lead to visual impairment due to diabetic retinopathy. In immortalized retinal pigment epithelial (RPE) cultures, high glucose concentrations are shown to lead to impairment in epithelial barrier properties. For the first time, the induced pluripotent stem-cell...

Purpose To study immunogenic properties of human embryonic stem cell–derived retinal pigment epithelium (hESC‐RPE) and to evaluate subretinal xenotransplantation of hESC‐RPE on porous polyethylene terephthalate (PET) in rabbits. Methods Human ESC‐RPE cells were characterized by morphology, transepithelial electrical resistance (TER), protein expre...

Retinal pigment epithelium (RPE) performs important functions for the maintenance of photoreceptors and vision. Malfunctions within the RPE are implicated in several retinal diseases for which transplantations of stem cell-derived RPE are promising treatment options. Their success, however, is largely dependent on the functionality of the transplan...

Supplementary Figure 1. Apical localization of Ca V 1.3 and Ca V 3.1 in hESC‐ and mouse RPE. Confocal microscopy xy‐maximum intensity projections and yz‐sections (apical side upwards, localization of the section highlighted with a white bar) show the immunolabeling of microvilli (ezrin, red) together with voltage‐gated Ca2+ channels A) CaV1.3 (gree...

Supplementary Figure 2. Changes in cell morphology during hESC‐RPE maturation. Reflection and bright field images of hESC‐RPE from post‐confluence day 1 to post‐confluence day 84 at four time points: A) day 1, B) day 6, C) day 31, and D) day 84 (cell line 08/017). Scale bars 5 μm.

Background: Wound healing of retinal pigment epithelium (RPE) is a complex process that may take place in common age-related macular degeneration eye disease. The purpose of this study was to evaluate whether wounding and wound healing has an effect on Ca2+ dynamics in human embryonic stem cell (hESC)-RPEs cultured different periods of time. Meth...

Pigmentation is a typical feature for retinal pigment epithelial cells (RPE), and each RPE cell represents individual characteristics. Although, type and intensity of pigmentation may vary, still most cells are pigmented to some extent. In this study, we set-up a standardized quantitative melanin content analysis for human embryonic stem cell (hESC...

Here we evaluated the effects of human retinal microvascular endothelial cells (hREC) on mature human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells. The hESC-RPE cells (Regea08/017, Regea08/023 or Regea11/013) and hREC (ACBRI 181) were co-cultured on opposite sides of transparent membranes for up to six weeks. Thereafter...

The impairment of autophagic and proteasomal cleansing together with changes in pigmentation has been documented in retinal pigment epithelial (RPE) cell degeneration. However, the function and co-operation of these mechanisms in melanosome-containing RPE cells is still unclear. We show that inhibition of proteasomal degradation with MG-132 or auto...

Degenerative retinal diseases are a leading cause of visual loss and irreversible blindness, particularly in the developed world. Retinal pigment cell (RPE) transplantation is nowadays considered the most promising therapeutic approach for certain retinal diseases, and the presence of a supportive scaffold has been considered essential to ensure th...

Human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cell therapies show tremendous potential for the treatment of retinal degenerative diseases. A tissue engineering approach, where cells are delivered to the subretinal space on a biodegradable carrier as a sheet, shows great promise for these RPE cell therapies. The aim of the...

Ca2+ is a second messenger controlling vital cellular processes, including cell maturation. Changes in Ca2+ signaling during maturation of human embryonic stem cell-derived retinal pigment epithelial cells (hESC-RPE) have not been assessed previously. The aim of this study was to investigate maturation-dependent changes in transient intracellular C...

Aims: A fast, non-invasive and observer-independent method to analyze the homogeneity and maturity of human pluripotent stem cell (hPSC) derived retinal pigment epithelial (RPE) cells is warranted to assess the suitability of hPSC-RPE cells for implantation or in vitro use. The aim of this work was to develop and validate methods to create ensembl...

Purpose: In several retinal complications, such as age-dependent macular degeneration (AMD), oxidative stress is increased and cytokine level is elevated. These are shown to alter the activation and expression of matrix metalloproteinase (MMP) both in human primary and immortalized retinal pigment epithelial (RPE) cells. However, the effects on hu...

Purpose The aim of this study was to investigate changes in spontaneous Ca ²⁺ activity and mechanically induced intercellular Ca ²⁺ communication in human embryonic stem cells‐derived retinal pigment epithelium ( hESC ‐ RPE ) during maturation. Methods In this study, we assessed Ca ²⁺ activity in hESC ‐ RPE cells cultured for 9 or 28 days. Ca ²⁺ i...

Human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells are currently undergoing clinical trials to treat retinal degenerative diseases. Transplantation of hESC-RPE cells in conjuction with a supportive biomaterial carrier hold great potential as a future treatment for retinal degeneration. However, there has been no such biod...

Computational models of calcium (Ca²⁺) signaling have been constructed for several cell types. There are, however, no such models for retinal pigment epithelium (RPE). Our aim was to construct a Ca²⁺ signaling model for RPE based on our experimental data of mechanically induced Ca²⁺ wave in the in vitro model of RPE, the ARPE-19 monolayer.We combin...

In in vitro live-cell imaging, it would be beneficial to grow and assess human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells on thin, transparent, rigid surfaces such as cover glasses. In this study, we assessed how the silanization of glass with 3-aminopropyltriethoxysilane (APTES), 3-(trimethoxysilyl)propyl methacrylate...

Objective Computational models of calcium (Ca 2+) signaling have been constructed for several cell types. There are, however, no such models for retinal pigment epithelium (RPE). Our aim was to construct a Ca 2+ signaling model for RPE based on our experimental data of mechanically induced Ca 2+ wave in the in vitro model of RPE, the ARPE-19 monola...

Bone morphogenetic protein 4 (BMP4) belongs to the transforming growth factor beta (TGF-beta) family of proteins. BMPs regulate cell proliferation, differentiation and motility, and have also been reported to be involved in cancer pathogenesis. We have previously shown that BMP4 reduces breast cancer cell proliferation through G1 cell cycle arrest...

Extracellular matrix (ECM) interactions play a vital role in cell morphology, migration, proliferation and differentiation of cells. We investigated the role of ECM proteins to the structure and function of human embryonic stem cell derived retinal pigment epithelial (hESC-RPE) cells during their differentiation and maturation from hESCs into RPE c...

Bone morphogenetic protein 4 (BMP4) is an extracellular signaling molecule that belongs to the bone morphogenetic protein (BMP) family, which in turn is part of the transforming growth factor β (TGF-β) -superfamily. BMPs regulate cell proliferation, differentiation and motility, and have also been shown to be involved in cancer pathogenesis. We hav...

Computational models of calcium (Ca2+) signaling have been constructed for several cell types. There are, however, no such models for retinal pigment epithelium (RPE). Our aim was to construct a Ca2+ signaling model for RPE based on our experimental data of mechanically induced Ca2+ wave in the in vitro model of RPE, the ARPE-19 monolayer. We combi...

Purpose: Aquaporins (AQPs), a family of transmembrane water channel proteins, are essential for allowing passive water transport through retinal pigmented epithelial (RPE) cells. Even though human native RPE cells and immortalized human RPEs have been shown to express AQPs, the expression of AQPs during the differentiation in stem cell-derived RPE...

Evidence is accumulating that ethanol and its oxidative metabolite, acetaldehyde, can disrupt intestinal epithelial integrity, an important factor contributing to ethanol-induced liver injury. However, ethanol can also be metabolized non-oxidatively generating phosphatidylethanol and fatty acid ethyl esters (FAEEs). This study aims to investigate t...

Ca(2+) signaling is vitally important in cellular physiological processes and various drugs also affect Ca(2+) signaling. Thus, knowledge of Ca(2+) dynamics is important toward understanding cell biology, as well as the development of drug-testing assays. ARPE-19 cells are widely used for modeling human retinal pigment epithelium functions and drug...

Purpose: Ocular surface reconstruction with cultivated oral mucosal epithelial transplantation technique is a viable treatment option for severe ocular surface injuries and diseases with limbal stem cell deficiency. Currently, this technique is based on utilization of xenogenic, allogenic or undefined components such as murine 3T3 feeders, serum a...

Purpose Our purpose is to construct mathematical models of functions of retinal pigment epithelium (RPE). Here we show the development and results of two models: 1) a compartmental model of Ca2+ dynamics 2) finite element model of epithelial transport and trans‐epithelial resistance. Methods A model of Ca2+ dynamics of APRE‐19 cells based on an exp...

Purpose Human pluripotent stem cell (hPSC) therapy is a potential approach for the replacement of degenerated retinal pigment epithelium (RPE) cells. For therapeutic use, safety and functionality of the RPE cells needs to be guaranteed. In addition to the basic cell and molecular biological characterizations it is vitally important to assess the di...

Intestinal barrier dysfunction and translocation of endotoxins are involved in the pathogenesis of alcoholic liver disease. Exposure to ethanol and its metabolite, acetaldehyde at relatively high concentrations have been shown to disrupt intestinal epithelial tight junctions in the conventional two dimensional cell culture models. The present study...

Retinal pigment epithelial (RPE) cells in the back of the eye nourish photoreceptor cells and form a selective barrier that influences drug transport from the blood to the photoreceptor cells. At the molecular level, ATP-dependent efflux transporters have a major role in drug delivery in human RPE. In this study, we assessed the relative expression...

Gene expression data with standard deviations (SD) and calculations of statistical significance (p). (DOC)

The barrier properties of epithelium are conventionally defined by transepithelial resistance (TER). TER provides information about the tightness of the epithelium. Electrical impedance spectroscopy (EIS) provides additional information regarding cell membrane properties, such as changes in electric capacitance and possible parallel or serial pathw...

The development and differentiation of stem cell-derived impermeable retinal pigment epithelium (RPE) with tight junctions (TJs) is a gradual process that is, at confluence, controlled by cell-to-cell contact. The objective of this study was to evaluate the use of electric impedance spectroscopy (EIS) to follow the maturation and development of bar...

Purpose Ocular surface reconstruction with cultivated oral mucosal epithelial transplantation technique has potential in the treatment of patients with severe ocular surface injuries. Currently, this technique is mainly based on utilization of xenogenic/allogenic components such as murine feeders, serum and amniotic membrane. The use of animal‐deri...

Purpose Stem cell therapy is a potential approach for the replacement of degenerated retinal pigment epithelial (RPE) cells essential for vision. For therapeutical use, the differentiation of RPE cells from human pluripotent stem cells (hPSC) should be performed in a culture environment fulfilling clinical quality requirements. For example the use...

Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) serve an unlimited source of cells capable of differentiating retinal pigment epithelial cells. These cells may be used in in vitro drug testing and in cell transplantation therapies. Small population of pluripotent hESCs and iPSCs spontaneously differentiate to RPE cells....

Three-dimensional reconstruction of a T84 sphere. Cells were cultured for 7 days in matrigel to form hollow spheres. Cells were processed for immunolabeling with antibodies against the adherens and tight junction proteins β-catenin (in red) and ZO-1 (in green), respectively. Confocal stacks (over sampled) were generated, deconvoluted, 3D reconstruc...

Caco-2 cells in 3D culture develop a polarized, yet heterogenous spheres. A) Low magnification phase contrast image of 7 days-old Caco-2 in 3D culture. B) gp135 (podocalyxin) and ZO-1 are exclusively expressed at the actin filament-rich luminal cell surface of the spheres, whereas the β-1 integrin receptor localizes to the basolateral domain of the...

IFNγ has no effect on 3D intestinal epithelial cell morphogenesis. A) Cells were plated in Matrigel in the presence or in the absence of IFNγ. The number of luminal spheres (expressed as percentage of all spheres) is depicted as function of time following cell plating. B) Cells were plated in Matrigel in the presence or in the absence of IFNγ The n...

The cytokines TNF (TNFSF2) and IFNγ are important mediators of inflammatory bowel diseases and contribute to enhanced intestinal epithelial permeability by stimulating apoptosis and/or disrupting tight junctions. Apoptosis and tight junctions are also important for epithelial tissue morphogenesis, but the effect of TNF and IFNγ on the process of in...

The production of functional retinal pigment epithelium (RPE) cells from human embryonic (hESCs) and human induced pluripotent stem cells (hiPSCs) in defined and xeno-free conditions is highly desirable, especially for their use in cell therapy for retinal diseases. In addition, differentiated RPE cells provide an individualized disease model and d...

Purpose Defined differentiation of functional RPE cells from human embryonic (hESC) or induced pluripotent stem cells (iPSC) is a prerequisite for their use in individualised disease modelling, drug discovery and transplantation for retinal diseases. In this study we report differentiation of RPE cells from hESC and iPSC in condition enabling easy...

Purpose Corneal epithelial reconstruction by using cultivated oral mucosal epithelial transplantation (COMET) technique is a viable treatment option for severe ocular surface disorders. So far this has been based on utilization of xenogenic/allogenic components such as 3T3 mouse feeder cells, serum or amniotic membrane. In this study, we present a...

In coeliac disease, small-bowel mucosal permeability is increased in response to gluten consumption. However, the signalling routes leading to such a barrier defect remain obscure. As the epidermal growth factor receptor (EGFR) pathway is up-regulated in untreated coeliac disease, and since this cascade has been related to epithelial hyperpermeabil...

Wheat gliadin induces severe intestinal symptoms and small-bowel mucosal damage in coeliac disease patients. At present, the only effective treatment for the disease is a strict life-long gluten-free diet. In this study we investigated whether probiotics Lactobacillus fermentum or Bifidobacterium lactis can inhibit the toxic effects of gliadin in i...

Coeliac disease is caused by dietary gluten, which triggers chronic inflammation of the small intestine in genetically predisposed individuals. In one quarter of the patients the disease manifests in the skin as dermatitis herpetiformis. Recently, a novel candidate gene, myosin IXB on chromosome 19p13, was shown to be associated with coeliac diseas...

In coeliac disease gluten induces an immunological reaction in genetically susceptible patients, and influences on epithelial cell proliferation and differentiation in the small-bowel mucosa. Our aim was to find novel genes which operate similarly in epithelial proliferation and differentiation in an epithelial cell differentiation model and in coe...

List of genes evincing a significant change in their mRNA expression in TGFβ-differentiated compared to T84 cells grown solely in collagen I gel (= TGFβ vs. control) and T84 cells differentiated by soluble factors secreted by mesenchymal cells compared to T84 cells grown solely in collagen I gel (= IMR-treated vs. control). Genes are sorted by func...

The scatter plot for the normalized data:T84 cells differentiated by soluble factors secreted by mesenchymal cells compared to undifferentiated T84 cells grown solely in collagen I gel.

Cluster analysis for unfiltered data. Clustering of genes was done with software programs Cluster and Treeview.

The scatter plot for the normalized data: TGFβ-differentiated compared to undifferentiated T84 cells grown solely in collagen I gel.

Cluster analysis, heatmap, for the data first normalized by the sum method and further normalized using a linear mixed model. The Clustering of genes was done with software programs Cluster and Treeview.

The TGFbeta1-induced signal transduction processes involved in growth and differentiation are only partly known. The three-dimensional epithelial differentiation model, in which T84 epithelial cells are induced to differentiate either with TGFbeta1 or IMR-90 mesenchymal cell-secreted soluble factors, is previously shown to model epithelial cell dif...

In coeliac disease in the jejunum of a genetically susceptible person wheat gliadin and related prolamins from rye and barley trigger an immunological reaction, which induces small-bowel mucosal transformations, villous atrophy and crypt hyperplasia. Though CD4+ specific T cells, intraepithelial lymphocytes, infiltrating plasma cells and autoantibo...

In this study we have used the Semliki forest virus expression system to transiently express chimeric proteins that contain transmembrane and cytoplasmic domains of the cation-independent mannose 6-phosphate receptor (CI-MPR) fused to chicken avidin. Immunofluorescence and electron microscopy studies showed that the chimeric protein with the entire...