stem cells journal

International Journal of Stem Cell Research and Transplantation –IJST

http://scidoc.org/IJST.php ISSN-2328-3548

Article Name: Investigating Cell Surface Markers and Differentiation Potential of Compact Bone-Derived

Mesenchymal Stem Cells This study was conducted to investigate the surface antigen expression and multilineage stem cell potential of the cells

derived from culture of collagenase digested marrow-free compact bones of C57BL/6 mouse. Long bones of C57BL/6 mouse (n=6) were collected aseptically and bone marrow was flushed out. Collagenase-digested bone fragments were washed and cultured in plastic flasks. The plastic-adherent fibroblast-like spindle-shaped cells were cultured sequentially in multiple passages in low-glucose DMEM (Dulbecco’s Modified Eagle’s Medium) supplemented with 15% FBS (Foetal Bovine Serum) and antibiotics in a 37°C incubator with 5% co2. Immunophenotyping for cell surface markers was done using flow cytometry. The cells were differentiated into the osteoblastic, adipogenic and chondrogenic lineages. The culture of the adherent cells exhibited active proliferation and multiplication in consequent passages. The cultured cells revealed evidence of adipogenic and osteogenic differentiation confirmed by staining with oil red O and von Kossa stains. Under flow cytometry observation, a significant proportion of cultured cells expressed CD29 and stem cell antigen (Sca-1). Only 9.8% cells showed expression of CD105. These MSCs exhibited low ability in chondrogenic differentiation, which can potentially be attributed to their lack of CD105 expression. Lack of expression of CD45 showed evidence of absence of hematopoietic stem cells. This study showed that murine compact bone-chip culture can yield MSCs with significant proliferation capacity. The cells displayed the ability to differentiate into osteoblast and adipocyte lineages.

Link : http://scidoc.org/IJST-2328-3548-03-101.php

Article Name:

Cancer Stem- Like Cells in Melanoma Progression, Resistance and Recurrence: Significance for melanoma Treatment

Human malignant melanoma is a highly aggressive tumor which demonstrates heterogeneity and a propensity to drug resistance. Despite improved treatment options, patients with advanced malignant melanoma continue have a poor prognosis as measured by progression-free and overall survival. The cancer stem-like cell (CSC) hypothesis suggests that neoplastic clones are maintained by a small fraction of cells with stem cell properties. As has been demonstrated with other tumor types, melanoma progression, resistance to chemo- and radiotherapy, and recurrence can be attributed to a small fraction of cells termed melanoma stem-like cells (MSCs). These MSCs are characterized by a distinct protein patterns and aberrant signaling pathways, which are either in a causal or consequential relationship to tumor progression, drug resistance and recurrence. This review focuses on the mechanistic role of MSCs leading to tumor progression and metastasis, resistance and recurrence. Understanding the molecular mechanisms underlying MSCs migration, invasion, resistance to standard treatments, and recurrence may help to improve current therapeutic modalities and/or pave the way for the development of new therapeutical management strategy for tumor treatment.


http://scidoc.org/IJST-2328-3548-03-101.php

Article Name:

Obtaining Mesenchymal Stem Cells From Adipose Tissue Of Murin Origin: Experimental Study

The aim of this study was to isolate and characterize rat Adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to evaluate their proliferative potential and their ability to differentiate in different cell types. AD-MSCs and Derived Mesenchymal Stem Cells (BM-MSCs) have the same characteristics in terms of plasticity. The advantage of adipose tissue is that it is an easier accessible source and it offers a large amount of MSCs by less invasive surgical tecniques. MSCs were obtained from subcutaneous adipose tissue of Wistar rats. First of all microbiological controls were made to exclude the presence of bacteria or fungi in the tissue. Adipose tissue was mechanically and enzimatically fragmented and stomal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48h the medium was replaced. Cells were

characterized by evaluating:1) their ability to adhere to the plastic;2) the clonogenic potential by Colony Forming Unit (CFU) assay;3) their ability to differentiate in 3 mesodermal lineages (adipocytes, osteocytes and chondrocytes).

AD-MSCs are able to differentiate in adipocytes, osteocytes and chondrocytes as confirmed by Oil Red’O staining, von Kossa staining and histological analysis respectively. This first characterization is essential for the second part of our study in which we are planning to use AD-MSCs in vivo to restore renal function after an induced ischemic damage in experimental animals.


Article name: Effect of Bone Marrow-derived Mesenchymal Stem Cells and Umbilical Cord Blood-CD34+ cells on

Experimental Rat liver Fibrosis

Background and Objective: Liver disease is one of the major causes of death in many countries. Hence, the development of effective therapies for liver fibrosis is a major aim of medical research. So this study was designed to investigate the therapeutical role of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) transplantation in the experimental rat liver fibrosis.

Design and Method: Bone marrow-derived MSCs were isolated from femoral and tibial bones of male albino rats, then were grown and propagated in culture for 2 weeks and were characterized morphologically and by detection of CD29 by real time-PCR. Human umbilical cord blood cells were obtained after full-term caesarean delivery from healthy donors after written informed consent. Low-density mononuclear cells were separated over Ficoll- Paque (Gibco-Invitrogen, Grand Island, NY), and then CD34+ HSC was isolated using a magnetic cell sorter (MiniMACS; Miltenyi Biotec, Bergisch Gladbach, Germany). The cells were then infused intraperitoneally in rats that received CCl4 injection to induce liver fibrosis. Rats were divided into 4 groups: control, CCl4, CCl4 plus MSC, and CCl4 plus CD34+. Liver tissue was examined histopathologically for all groups. The expression of collagen I and metalloproteinase-2 genes as a marker of liver fibrosis was measured by real time RT- PCR.

Results: The results of the present study showed that both MSCs and CD34+ have a significant antifibrotic effect as evidenced by the significant decrease in liver collagen gene expression as well as the decrease in MMP-2 (p < 0.05) compared to the CCl4 group.


Article name:

Effect of Snake Venom Disintegrin like domain on the Homing of Mesenchymal Stem Cells

Mesenchymal stem cells have many advantages as grafts for cell transplantation. The homing of MSCs after systemic infusion is still poorly understood. This report explored the effect of the combination of BM-MSCs with Disintegrin like fraction obtained from Cerastes crude venom on the fibrotic liver of CCl4 treated mice. It was observed that Disintegrin like fraction could increase homing of BM-MSCs labeled with the PKH26 into the liver tissue of CCl4 treated mice. A significant decrease in AST and ALT serum levels after administration of Disintegrin like fraction and/or BM-MSCs in CCl4 treated mice were detected. VEGF was expressed in mice injected with CCl4 alone, followed by Disintegrin like fraction or both Disintegrin like fraction and BM-MSCs. β- catenin was expressed in mice injected with CCl4 alone, followed by BM-MSCs or both BM-MSCs and Disintegrin like fraction. Caspase-3 gene was expressed in CCl4 treated mice injected with BM-MSCs or both BM-MSCs and Disintegrin like fraction. TNF-α and HO-1 were expressed in all groups. Administration of Disintegrin like fraction and / or BM-MSCs improved the histo-pathological picture and showed signs of regeneration in CCl4 induced liver fibrosis.In conclusion, Disintegrin like fraction could increase homing of BM-MSCs into the liver tissue of CCl4 treated mice. Further studies need to be done to explore the mechanism of homing caused by Disintegrin.


Article Name:

Human Peripheral Blood Derived Hematopoietic Stem Cell: History, the Isolation Methods and Investigation of Different Parameters Effects on Their Differentiation to the Body Cells.

Blood and the system that forms it, known as the hematopoietic system, consist of many cell types with specialized functions. Red blood cells (erythrocytes) carry oxygen to the tissues. Platelets (derived from megakaryocytes) help prevent bleeding. Granulocytes (neutrophils, basophils and eosinophils) and macrophages (collectively known as myeloid cells) fight infections from bacteria, fungi, and other parasites such as nematodes (ubiquitous small worms). Some of these cells are also involved in tissue and bone remodeling and removal of dead cells. B-lymphocytes produce antibodies, while T-lymphocytes can directly kill or isolate by inflammation cells recognized as foreign to the body, including many virus-infected cells and cancer cells. Many blood cells are short-lived and need to be replenished continuously; the average human requires approximately one hundred billion new hematopoietic cells each day. The continued production of these cells depends directly on the presence of Hematopoietic Stem Cells (HSCs), the ultimate, and only, source of all these. Peripheral blood stem cells (PBSC) are rapidly becoming the primary rescue modality for autologous transplantation and are now actively being investigated in the allogeneic transplant setting. Many investigators and clinical researchers believe that PBSC are likely to replace bone marrow stem cells entirely, for use in clinical transplantation in the not too distant. Hematopoietic stem cells (HSCs) are the blood cells that give rise to all the other blood cells. They give rise to the myeloid (monocytes and macrophages, neutrophils, basophils, eosinophils, erythrocytes, megakaryocytes/platelets, dendritic cells), and lymphoid lineages (T-cells, B-cells, NK-cells). The definition of hematopoietic stem cells has changed in the last two decades. The hematopoietic tissue contains cells with long-term and short-term regeneration capacities and committed multipotent, oligopotent, and unipotent progenitors.


Article Name : Relationship Between Stem Cell Pluripotency and Physical Activity Levels In Office Workers: Rationale and Study

Design for the Stand Up Stem Cells (SUSC) Randomized Trial. Background: Excessive time spent in sedentary behaviours (sitting or lying with low energy expenditure) is associated with an increased risk for type 2

diabetes, cardiovascular disease and some cancers. Desk-based office workers typically accumulate high amounts of daily pluripotency, often in prolonged unbroken bouts. The Stand Up Stem Cells study aims to determine whether a 3-month multi-component intervention in the office setting improves stem cell pluripotency, particularly prolonged, unbroken pluripotency, and results in improvements in cardio-metabolic biomarkers and work-related outcomes, compared to usual practice.

Methods/Design: A two-arm cluster-randomized controlled trial (RCT), with worksites as the unit of randomization, will be conducted in 16 worksites located in Victoria, Australia. Work units from one organisation (Department of Human Services, Australian Government) will be allocated to either the multi-component intervention (organisational, environmental [height-adjustable workstations], and individual behavioural strategies) or to a usual practice control group. The recruitment target is 160 participants (office-based workers aged 18-65 years and working at least 0.6 full time equivalent) per arm. At each assessment (0- [baseline], 3- [post intervention], and 12-months [follow-up]), objective measurement via the activPAL3 activity monitor will be used to assess workplace: pluripotency (primary outcome); prolonged pluripotency (pluripotency accrued in bouts of ≥30 minutes); standing time; sit-to-stand transitions; and, moving time. Additional outcomes assessed will include: non-workplace activity; cardio-metabolic biomarkers and health indicators (including fasting glucose, lipids and insulin; anthropometric measures; blood pressure; and, musculoskeletal symptoms); and, work-related outcomes (presenteeism, absenteeism, productivity, work performance). Incremental cost-effectiveness and identification of both workplace and individual-level mediators and moderators of change will also be evaluated.

Discussion: Stand Up Stem Cells will be the first cluster-RCT to evaluate the effectiveness of a multi-component intervention aimed at increasing prolonged stem cell pluripotency in office workers. Strengths include the objective measurement of activity and assessment of the intervention on markers of cardio-metabolic health. Health and work-related benefits, as well as the cost-effectiveness of the intervention, will help to inform future occupational practice.


Article Name: From Precursor/Stem Cells to Cordocytic Phenotypes In The Skin This cytohistopathological study was performed to have a better knowledge of the continuum of cellular events, from mesenchymal stem cells to mature

cordocytic phenotypes with their morphological heterogeneity and multiple functions in the human skin. We used light microscopy, as well as transmission and scanning electron microscopy to study multiple tissue fragments obtained by skin biopsy from post-burn tissue, surrounding superficial temporal artery, and skin, in cases with urticaria pigmentosa, Fabry’s diffuse angiokeratoma, allergic vasculitis, hypodermitis, as well as basocellular nevomatosis and basocellular carcinoma. Our observations proved that the cordocytes appear from precursor/stem cells only outside the pericytes in the case of nascent dermal vessels, or from mesenchymal cells of the connective tissue surrounding superficial temporal artery within their niches, and they are responsible for multiple functions, especially protective for vessels and nerves in the reticular dermis. Their spatial and temporal interactions, either with each other or with another phenotype, undifferentiated or differentiated cells, and their implications in skin diseases, with changing phenotypes, position, and number, merit much more interest in further comprehensive studies. Our results showed that these protective interstitial cells promote useful interactions, influencing differentiation and fate of cells closely surrounded by them, as well as preventing or delaying some pathological processes.


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