Characterization Of Mesenchymal Stem Cells In Mucolipidosis Type Ii (I-Cell Disease)

Mucolipidosis type II (ML-II, I-cell disease) is a fatal inherited lysosomal storage disease caused by a deficiency of the enzyme N-acetylglucosamine-1-phosphotransferase. A characteristic skeletal phenotype is one of the many clinical manifestations of ML-II. Since the mechanisms underlying these skeletal defects in ML-II are not completely understood, we hypothesized that a defect in osteogenic differentiation of ML-II bone marrow mesenchymal stem cells (BM-MSCs) might be responsible for this skeletal phenotype. Here, we assessed and characterized the cellular phenotype of BM-MSCs from a ML-II patient before (BBMT) and after BM transplantation (ABMT), and we compared the results with BM-MSCs from a carrier and a healthy donor. Morphologically, we did not observe differences in ML-II BBMT and ABMT or carrier MSCs in terms of size or granularity. Osteogenic differentiation was not markedly affected by disease or carrier status. Adipogenic differentiation was increased in BBMT ML-II MSCs, but chondrogenic differentiation was decreased in both BBMT and ABMT ML-II MSCs. Immunophenotypically no significant differences were observed between the samples. Interestingly, the proliferative capacity of BBMT and ABMT ML-II MSCs was increased in comparison to MSCs from age-matched healthy donors. These data suggest that MSCs are not likely to cause the skeletal phenotype observed in ML-II, but they may contribute to the pathogenesis of ML-II as a result of lysosomal storage-induced pathology.PubMedWoSScopu

Osteopetrotic Induced Pluripotent Stem Cells Derived From Patients With Different Disease-Associated Mutations By Non-Integrating Reprogramming Methods

Background Autosomal recessive osteopetrosis is a genetically and phenotypically heterogeneous disease, caused by defects in osteoclast formation and function. The only available treatment is allogeneic stem cell transplantation that has still high morbidity and mortality. The goal of the present study was to generate iPSCs from bone marrow-derived MSCs of osteopetrosis patients with three most common mutations by using two different integration-free gene transfer methods and compare their efficiencies. The secondary objective was to select the most appropriate integration-free production method for our institutional iPSC bank using this rare disease as a prototype. Methods Two different integration-free gene transfer methods (episomal and Sendai viral vectors) were tested and compared on the same set of patient samples exhibiting three different mutations associated with osteopetrosis. Generated iPSCs were characterized by standard assays, including immunophenotyping, immunocytochemistry, RT-PCR, embryoid body, and teratoma assays. Karyotype analyses were performed to evaluate genetic stability. Results iPSC lines exhibiting typical ESC-like colony morphology were shown to express pluripotency markers by immunofluorescence staining. Over 90% of the cells were found positive for SSEA-4 and OCT3/4 and negative/weak positive for CD29 by flow cytometry. Immunohistochemical staining of teratoma and spontaneously differentiated embryoid body sections confirmed their trilineage differentiation potential. All iPSC lines expressed pluripotency-related genes. Karyotype analyses were found normal. Direct sequencing of PCR-amplified DNA showed that disease-related mutations were retained in the patient-specific iPSCs. Conclusion Generation of iPSC using SeV and episomal DNA vectors have several advantages over other methods like the ease of production, reliability, high efficiency, and safety, which is required for translational research. Furthermore, owing to the pluripotency and self-renewal capacity, patient-specific iPSCs seem to be ideal cell source for the modeling of a rare genetic bone disease like osteopetrosis to identify osteoclast defects, leading to clinical heterogeneity in osteopetrosis patients, especially among those with different mutations in the same gene. Electronic supplementary material The online version of this article (10.1186/s13287-019-1316-8) contains supplementary material, which is available to authorized users.PubMedWoSScopu

Hematopoetik Kök Hücre Hibernasyonunda Nöropeptid Y (NPY)´nin Rolünün İncelenmesi

Hematopoetik kök hücreler (HKH), kök hücre nişi olarak da adlandırılan özel bir mikroçevrede bulunur. Bu mikrocevrede mezenkimal kök hucreler (MKH), endotelyal hücreler (EH), retiküler hücreler bulunur. Perivasküler mikroortamda bulunan EH, MKH, makrofajlar ve HKH yaşamını ve kemik iliği rejenerasyonunu düzenler.Nörotransmitter nöropeptit Y (NPY), makrofajlardan ve osteoblastlardan salınır. NPY, immün ve kemik hücre homeostazını veya bu yapıların vasküler yeniden yapılanmasını, kemik iliği hücrelerinde ifade edilen Y reseptörüyle düzenlerler. Hibernasyonda HKH'ler sıklıkla trabeküler endosteuma yakın bulunurlar. İhtiyaç duyulduğunda bu hücreler çoğalır ve farklılaşır, stres ve yaralanmalarda salgılanan büyüme faktörleri ile hücre döngüsüne girerler. NPY eksikliğinde, HKH’lerin sayıları düşmekte ve kemik iliği rejenerasyonu bozulmaktadır. Önerilen projenin amacı, NPY ve NPY reseptör sinyal yolağı etkisiyle HKH'lerin uyku durumuna geri dönmesi ve HKH havuz boyutu ile kemik iliği fonksiyonunun korunmasında sessizliğin rolünün ortaya çıkmasına ilişkin anahtar bir mekanizmanın belirlenmesidir

Hyperthyroidism After Allogeneic Hematopoietic Stem Cell Transplantation: A Report Of Four Cases

Hematopoietic stem cell transplantation (HSCT) is the only curative treatment for many hematological disorders, primary immunodeficiencies, and metabolic disorders. Thyroid dysfunction is one of the frequently seen complications of HSCT. However, hyperthyroidism due to Graves’ disease, autoimmune thyroiditis, and thyrotoxicosis are rare. Herein, we report a series of 4 patients who were euthyroid before HSCT but developed hyperthyroidism (3 of them developed autoimmune thyroid disease) after transplantation.PubMedWoSScopu

Venous Thromboembolism after Allogeneic Pediatric Hematopoietic Stem Cell Transplantation: A Single-Center Study

Objective: Venous thromboembolism (VTE) in children who undergo hematopoietic stem cell transplantation (HSCT) has high morbidity. The aim of this study is to assess the incidence of VTE in allogeneic pediatric HSCT recipients and the contribution of pretransplant prothrombotic risk factors to thrombosis. Materials and Methods: We retrospectively evaluated 92 patients between April 2010 and November 2012 undergoing allogeneic HSCT who had completed 100 days post-HSCT. Before HSCT, coagulation profiles; acquired and inherited prothrombotic risk factors including FV G1691A (factor V Leiden), prothrombin G20210A, methylenetetrahydrofolate reductase (MTHFR) C677T, and MTHFR A1298C mutations; and serum homocysteine and lipoprotein (a), plasma antithrombin III, protein C, and protein S levels were obtained from all patients. Results: In the screening of thrombophilia, 8 patients (9%) were heterozygous for factor V Leiden, 5 (6%) were homozygous for MTHFR 677TT, 12 (14%) were homozygous for MTHFR 1298CC, and 2 (2%) were heterozygous for prothrombin G20210A mutation. We observed VTE in 5 patients (5.4%); a prothrombotic risk factor was found in 3 out of these 5 patients, while 4 out of 5 patients had central venous catheters. It was determined there was no significant relationship between VTE and inherited prothrombotic risk factors. Conclusion: VTE after HSCT seems to be a low-frequency event that may be due to low-dose, low-molecular-weight heparin prophylaxis, and the role of inherited prothrombotic risk factors cannot be entirely excluded without a prospective study

Thrombotic Microangiopathy in Allogeneic Stem Cell Transplantation in Childhood

OBJECTIVES: We define the incidence, risk factors, and mortality rates for the occurrence of thrombotic microangiopathy in 50 children who underwent transplants between January 2006 and June 2008 at 2 Turkish pediatric centers. MATERIALS AND METHODS: The diagnosis of thrombotic microangiopathy was done according to the reports of International Working Group in 2007. RESULTS: Fifty patients (27 male and 23 female; age range, 3 months to 18 years) were included. Patients with malignant and nonmalignant diseases were 13 (26%) and 37 (74%). Myeloablative and nonmyeloablative conditioning regimens were used in 29 (58%) and 21 patients (42%). Bone marrow was used as the source of stem cells in 32 patients (62%) and peripheral blood was used in 18 patients (36%). Thrombotic microangiopathy was seen in 3 of 50 cases (6%). Thrombotic microangiopathy developed in 3 of 18 patients in whom peripheral blood was used as the source of stem cells while none of 32 patients who had bone marrow as the source developed thrombotic microangiopathy (P < .05). CONCLUSIONS: Using peripheral blood as a source of stem cells is a risk factor for development of thrombotic microangiopathy.Wo

Effect of Ocimum basilicum on mesenchymal stem cell proliferation and differentiation: Does the effect change according to niches?

It is a big issue that reduced bone density and large fractıres in dentistry and orthopedics. Side effects caused by synthetic drugs lead to medical and ethical problems. Thus, plants and medicinal plant research take attention. Aim of this preliminary in vitro study is to investigate the effect of Ocimum basilicum extract on dental pulp (DP) and bone marrow (BM) derived mesenchymal stem cell (MSC) proliferation, osteogenic differentiation and immunological response to TNF-α. Human dental pulp tissue was obtained from patients (15-20 years of age) who were undergoing extraction of third molars for orthodontic reasons at the Department of Oral and Maxillofacial Surgery, University of Gazi University*. xCELLigence system was used to determine prolfieration of DP- and BM-MSCs. Adipogenic and osteogenic differentiation was shown and calcium concentration, osteocalcin and osteonectin levels were examined. Inflammatory environment was mimiced through TNF-α stimulation and IL-6 and IL-10 levels were defined by ELISA. Doubling time mwith O. basilicum was found in DP- MSCs (38 h) and BM-MSCs (76 h). IC50 value was shown as 148 µg/mL in DP-MSCs and 178 µg/mL in BM-MSCs. Calcium concentration of BM-MSCs was found decreased in O. basilicum treated groups. Level of ostoenectin was reduced in O. basilicum treated cells suggesting that the Extract accelerated the osteogenic differentiation. We suggest that O. basilicum could be a smart ostoeinductive agent where BM-MSCs should be investigated further. Rich flora of Turkey is an opportunity for us and encourangement can easily give inside to medicinal plant investigations. *B.30.2.GÜN.0.20-122 Ethics Committee Repor

Diagnosis: Melanoderma After Hematopoietic Stem Cell Transplantation

PubMedScopu

Automated washing of long-term cryopreserved peripheral blood stem cells promotes cell viability and preserves CD34+ cell numbers

Peripheral blood stem cell (PBSC) transplantation has become an established treatment option for a range of malignant and inherited diseases. PBSCs are usually cryopreserved in the presence of dimethyl sulfoxide (DMSO) and stored in liquid nitrogen. However, cryopreservation and thawing of PBSCs may affect cell viability, resulting in delayed engraftment and other risks related to low stem cell numbers [1]. DMSO is commonly used at a concentration of 10% during freezing of PBSCs. However, DMSO itself is toxic and lowering doses of DMSO may have a favorable effect on hematopoietic recovery after transplantation [2, 3]. Generally, PBSCs are thawed and infused without removal of DMSO. This has been associated with a wide range of adverse effects (AEs), ranging from minor to severe life-threatening events [4, 5]. Although not all toxic events can be contributed to DMSO, grafts containing lower concentrations of DMSO typically display a reduced incidence of AEs [5, 6]. To decrease AEs and improve graft quality after thawing, it has been suggested to remove DMSO prior to transplantation [4, 7] using different washing systems [8, 9]. Here, we aimed to remove DMSO from long-term cryopreserved PBSCs using an automated, fully closed system and assessed effects on CD34+ stem cells, viability, and colony-forming capacity