Odlade mesenkymala stamcellers användning vid skador på perifera och centrala nervsystemet

Bone marrow-derived mesenchymal stem cells (MSC) have been shown to provide neuroprotection after transplantation into the injured nervous system. The present thesis investigates whether adult human and rat MSC differentiated along a Schwann cell lineage could increase their expression of neurotrophic factors and promote regeneration after transplantation into the injured peripheral nerve and spinal cord. Human and rat mesenchymal stem cells (hMSC and rMSC) expressed characteristic stem cell surface markers, mRNA transcripts for different neurotrophic factors and demonstrated multi-lineage differentiation potential. Following treatment with a cocktail of growth factors, the hMSC and rMSC expressed typical Schwann cells markers at both the transcriptional and translational level and significantly increased production of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF). Age and time in culture are of relevance for clinical settings and growth-promoting effects of hMSC from young donors (16-18 years) and old donors (67-75 years) were compared. Undifferentiated hMSC from both young and old donors increased total neurite length of cultured dorsal root ganglion (DRG) neurons. Differentiation of hMSC from the young donors, but not the eldery donors, further enhanced the neurite outgrowth. Undifferentiated hMSC were cultured for eleven weeks in order to examine the effect of in vitro expansion time on neurite outgrowth. hMSC from the young donors maintained their proliferation rate and their ability to enhance neurite outgrowth from DRG neurons. Using a sciatic nerve injury model, a 10mm gap was bridged with either an empty tubular fibrin glue conduit, or conduits containing hMSC, with and without cyclosporine treatment. Cells were labeled with PKH26 prior to transplantation. At 3 weeks after injury the conduits with cells and immunosuppression increased regeneration compared with an empty conduit. PKH26 labeled human cells survived in the rat model and the inflammatory reaction could be suppressed by cyclosporine. After cervical C4 hemisection, BrdU/GFP-labeled rMSC were injected into the lateral funiculus rostral and caudal to the spinal cord lesion site. Spinal cords were analyzed 2-8 weeks after transplantation. Transplanted MSC remained at the injection sites and in the trauma zone for several weeks and were often associated with numerous neurofilament-positive axons. Transplanted rMSC induced up-regulation of vascular endothelial growth factor in spinal cord tissue rostral to the injury site, but did not affect expression of brain-derived neurotrophic factor. Although rMSC provided neuroprotection for rubrospinal neurons and significantly attenuated astroglial and microglial reaction, cell transplantation caused aberrant sprouting of calcitonin gene-related peptide immunostained sensory axons in the dorsal horn. In summary these results demonstrate that both rat and human MSC can be differentiated towards the glial cell lineage, and show functional characteristics similar to Schwann cells. hMSC from the young donors represent a more favorable source for neurotransplantation since they maintain proliferation rate and preserve their growth-promoting effects in long-term cultures. The data also suggest that differentiated MSC increase expression of neurotrophic factors and support regeneration after peripheral nerve and spinal cord injury

Evaluation of growth, stemness, and angiogenic properties of dental pulp stem cells cultured in cGMP xeno-/serum-free medium

This study was aimed to investigate the effects of cGMP xeno-/serum-free medium (XSF, Irvine Scientific) on the properties of human dental pulp stem cells (DPSCs). DPSCs, from passage 2, were cultured in XSF or fetal bovine serum (FBS)-supplemented medium, and sub-cultured up to passage 8. Cumulative population doublings (PDs) and the number of colony-forming-units (CFUs) were determined. qRT-PCR, ELISA, and in vitro assays were used to assess angiogenic capacity. Flow cytometry was used to measure CD73, CD90, and CD105 expression. Differentiation into osteo-, adipo-, and chondrogenic cell lineages was performed. DPSCs showed more elongated morphology, a reduced rate of proliferation at later passages, and lower CFU counts in XSF compared with FBS. Expression of angiogenic factors at the gene and protein levels varied in the two media and with passage number, but cells grown in XSF had more in vitro angiogenic activity. The majority of early and late passage DPSCs cultured in XSF expressed CD73 and CD90. In contrast, the percentage of CD105 positive DPSCs in XSF medium was significantly lower with increased passage whereas the majority of cells cultured in FBS were CD105 positive. Switching XSF-cultured DPSCs to medium supplemented with human serum restored the expression of CD105. The tri-lineage differentiation of DPSCs cultured under XSF and FBS conditions was similar. We showed that despite reduced CD105 expression levels, DPSCs expanded in XSF medium maintained a functional MSC phenotype. Furthermore, restoration of CD105 expression is likely to occur upon in vivo transplantation, when cells are exposed to human serum

Transformations of capitals: A case study of the museum of Imants Ziedonis

Bakalaura darbs “Kapitālu transformēšana: I. Ziedoņa topošā muzeja gadījums” ir pētījums par muzeja izveides procesu, apskatot kādi aspekti iezīmējas kā būtiski muzeja institūcijas izveidē un kādas kapitālu transformācijas var tikt vērojamas tās laikā. Pētījuma mērķis ir noskaidrot kā caur praksēm un zināšanu plūsmām notiek muzeja intstitūcijas izveides process, konkrētajā gadījumā - I. Ziedoņa topošais muzejs, apskatot to kapitālu transformāciju kontekstā. Pētījuma rezultāti ļauj spriest par I. Ziedoņa topošā muzeja izveides procesu kā zināšanu apmaiņas un stratēģisku prakšu kopumu, kurā tiek transformēti ekonomikas, kultūras, sociālais un simboliskais kapitāls.Bachelor’s thesis “Transformations of Capitals: A case study of the museum of Imants Ziedonis” is a research about the process of establishment of a museum, clarifying the transformations of capitals and aspects of the establishment of a museum. The aim of this research was to find out how through practices and knowledge the process of establishment of a museum is made, considering it through the transformations of capitals. The results of this research allow to reason about the process of establishment of the museum of Imants Ziedonis as an exchange of knowledge and strategic practices, in which economic, social, cultural and symbolic capital is being transformed

Water jet-assisted lipoaspiration and Sepax cell separation system for the isolation of adipose stem cells with high adipogenic potential

Introduction: Water jet-assisted liposuction has gained popularity due to favourable fat grafting outcomes. In this study, we compared stem cells obtained from fat isolated with manual or the water jet-assisted procedure. Methods: Liposuction of abdominal fat was performed using the two methods on each donor (n = 10). Aspirate samples were collagenase digested and the isolated cells seeded in vitro prior to proliferation, adipogenic differentiation and angiogenic activity analyses. Results: Cells from either procedure proliferated at similar rates and exhibited a similar colony-forming ability. The cells expressed stem cell markers CD73, CD90 and CD105. In the water jet cell preparations, there were higher numbers of cells expressing CD146. Robust adipogenic differentiation was observed in cultures expanded from both manual and water jet lipoaspirates. Gene analysis showed higher expression of the adipocyte markers aP2 and GLUT4 in the adipocyte-differentiated water jet cell preparations, and ELISA indicated increased secretion of adiponectin from these cells. Both cell groups expressed vasculogenic factors and the water jet cells promoted the highest levels of in vitro angiogenesis. Given these positive results, we further characterised the water jet cells when prepared using an automated closed cell processing unit, the Sepax-2 system (Cytiva). The growth and stem cell properties of the Sepax-processed cells were similar to the standard centrifugation protocol, but there was evidence for greater adipogenic differentiation in the Sepax-processed cells. Conclusions: Water jet lipoaspirates yield cells with high adipogenic potential and angiogenic activity, which may be beneficial for use in cell-assisted lipotransfers

Change in undifferentiated stem cell morphology and proliferation after time in culture.

<p>(<b>A</b>) Undifferentiated MSC showed spindle shaped morphology at T2 but cells from old donors show a bigger more flattered shape at T12. <b>(B</b>) uMSC were cultured for nine weeks and passaged and counted on a weekly basis. Cumulative population doublings were calculated and plotted against time.</p

Long-Term Effects of Fibrin Conduit with Human Mesenchymal Stem Cells and Immunosuppression after Peripheral Nerve Repair in a Xenogenic Model

Introduction: Previously we showed that a fibrin glue conduit with human mesenchymal stem cells (hMSCs) and cyclosporine A (CsA) enhanced early nerve regeneration. In this study long term effects of this conduit are investigated. Methods: In a rat model, the sciatic nerve was repaired with fibrin conduit containing fibrin matrix, fibrin conduit containing fibrin matrix with CsA treatment and fibrin conduit containing fibrin matrix with hMSCs and CsA treatment, and also with nerve graft as control. Results: At 12 weeks 34% of motoneurons of the control group regenerated axons through the fibrin conduit. CsA treatment alone or with hMSCs resulted in axon regeneration of 67% and 64% motoneurons respectively. The gastrocnemius muscle weight was reduced in the conduit with fibrin matrix. The treatment with CsA or CsA with hMSCs induced recovery of the muscle weight and size of fast type fibers towards the levels of the nerve graft group. Discussion: The transplantation of hMSCs for peripheral nerve injury should be optimized to demonstrate their beneficial effects. The CsA may have its own effect on nerve regeneration

Mesenchymal stem cells enhance neurite outgrowth from the human SH-SY5Y neuronal cell line.

<p>(<b>A</b>) Immunocytochemical staining for βIII tubulin (Alexa Fluor 488) visualized neurite outgrowth (highlighted with arrows) from SH-SY5Y cells following 24 h co-culture with medium only controls (α-MEM and diff α-MEM) and undifferentiated MSC (uMSC) and differentiated MSC (dMSC) from young and old donors seeded in tissue culture inserts above the neurons. (<b>B</b>) Quantitative analysis of total neurite outgrowth in the co-cultures. ***P<0.001 significantly different to respective media only controls; *P<0.05 significantly different young dMSC versus old dMSC; ns not significantly different to respective media only controls.</p

Late passage mesenchymal stem cells enhance neurite outgrowth of co-cultured DRG neurons.

<p>(<b>A</b>) Immunocytochemical staining for βIII tubulin (Alexa Fluor 488) visualized neurite outgrowth from DRG following 24 h co-culture with medium only controls (α-MEM and diff α-MEM) and undifferentiated MSC (uMSC) and differentiated MSC (dMSC) from young and old donors (T12) seeded in tissue culture inserts above the neurons. (<b>B</b>) Quantitative analysis of total neurite outgrowth in the co-cultures. ***P<0.001 significantly different to respective media only controls. n.s not significantly different (α-MEM and diff α-MEM versus DRG alone and dMSC versus uMSC old donors).</p