10th World Congress and Expo on Cell & Stem Cell Research March 19-21, 2018 New York, USA

Mon-Juan Lee has completed her MS degree in Chemical Engineering in 2000 and PhD degree in Life Science in 2006 from National Tsing Hua University, Hsinchu, Taiwan. She is currently an Associate Professor of the Department of Bioscience Technology with joint affiliation in the Graduate Institute of Medical Sciences at Chang Jung Christian University, Tainan, Taiwan. Her research interests include stem cell and bone biology, nanobiotechnology, as well as LC-based biomedical applications.

Sclerostin, a secreted glycoprotein expressed predominantly in osteocytes that inhibits bone formation by antagonizing the Wnt/β-catenin signaling pathway, is considered a new target for the treatment of osteoporosis. We devised a novel aptamer-based competitive drug screening platform for small-molecule sclerostin inhibitors from libraries of biologically active chemical compounds to facilitate drug repurposing and drug discovery for osteoporosis. Baicalein, a flavonoid and alizarin, an anthraquinone, are two of the potential sclerostin inhibitors obtained from the aptamer-based competitive assay and verified by an antibody-based competitive assay. The osteogenic potential of baicalein and alizarin was assessed by their capability to suppress the protein expression of sclerostin and to stimulate alkaline phosphatase activity. When mouse bone cell line IDG-SW3 was treated with baicalein or alizarin prepared in osteogenic induction medium, protein expression of sclerostin decreased compared to cells incubated in proliferation medium and osteogenic induction medium. In the presence of baicalein or alizarin, osteogenic differentiation was accelerated in human fetal osteoblasts hFOB1.19, in which higher alkaline phosphatase activity was detected than the control and cells treated only with osteogenic induction medium. Potential small-molecule drug candidates obtained in this study is expected to provide new therapeutics, as well as more insights into the structure–activity relationship of sclerostin inhibitors for further rational drug design. The common features in the chemical structure of baicalein and alizarin and their osteogenic potential demonstrated in this study suggest that flavonoids and anthraquinones may serve as lead compounds of small-molecule inhibitors of sclerostin for the development of bonestimulating new drugs.

Milla Lampinen has received her PhD from University of Helsinki in 2003 from Prof. Kari Keinänen’s lab, studying glutamate receptor ligand binding determinants. She has completed her Post-doctoral training from Prof. Anu Wartiovaara’s group studying mitochondrial disease and at Finnish Red Cross Research and development unit, studying stem cells and product applications thereof. She is currently working on stem cell behavior and cardiac stem cell therapy studies in Dr. Esko Kankuri’s group at University of Helsinki.

The loss of cardiac myocytes and the nonreversible decrease in cardiac output are the major challenges of ischemic myocardial injury (MI) and chronic heart failure (HF). Recent studies indicate that cardiac-derived cells offers new potential to repair myocardium with sustained functional benefit. We present here a novel, cost-effective cell-patch method, to non-invasively harvest AA cell-micrograft from surgical left-over tissue of CABG and to deliver them within scaffold to provide easy epicardial targeting and intraoperative treatment of HF. We applied the cellular therapy protocol both in mouse acute MI-model and in human safety-feasibility trial. In mouse, allogeneic mouse AA-cells containing scaffold are place on infarction area. For eight weeks echocardiographic follow-up, the cell-on-scaffold treatment showed statistically higher functional recovery in under curve-analysis of left ventricle ejection fraction (LVEF) than the empty-scaffold and non-treated infarction. Also, remodeling was significantly reduced by scaffold+/-cells, measured by the larger relative wall thickness of the stained collagen in nontreated group. The highest tissue expression of HF identifier, natriuretic peptide, in infarction group further suggests the AA-cell therapy’s beneficial effect on heart survival, while empty-scaffold may also provide structural support to ventricle wall. In clinical safety trial, six patients undergoing elective CABG are treated with diameter of 2.5cm cell-on-scaffold. In follow-up of first patient at three months, magnetic resonance imaging revealed good performance of the heart and visible thickened ventricle wall in comparison to pre-operative analysis. The patient’s generally good recovery from the operation further suggests that the protocol is feasible for operation room use and the cell-micrograft therapy is safe.

Shirin Nouraein has completed her MS degree in Biomedical/Tissue Engineering from University of Tehran and had cooperation with skin team at University of Tehran. Currently, she is working at Royan Instituate of Stem Cell and Medicine Regenerative as a Research Assistance.

Recently, protocols based on stem cell therapies have helped to understand long-sight goals in transplantation. Many substantial progresses have been made over the years using stem cells for improving successful transplants and decrease the rate and severity of graft rejection, modulation of regulatory cell types, accelerate and improve damaged tissue regeneration and induction of immune tolerance and immunosuppression. There is no doubt that the use of mesenchymal stem cells in transplantation fields had a positive effect on having a better outcome. The main goal of this experiment was to isolate and purificate mesenchymal stem cells from mice and administration of MSCs to decreasing the rate and severity of skin allograft rejection in mice. Animals divided into three individual groups which had a single dose of systemic injections of MSCs in different times of allotransplantation of C57BL/6 mice skin graft to Balb/C recipient. Our results have shown that autologous MSCs can extend the time until skin graft rejection and its best to administer MSCs after transplantation. This result is probably because of migration of MSCs to graft site due to local inflammation reactions that has already existed in recipient body because of transplantation. So, injecting cells after transplantation has a better effect. But, there is still a lot of variables to study and proof this hypothesis. In the other hand, MSCs from donor can also can extend the graft survival, even though they are allogeneic to recipient’s body and activate immune system. So the mesenchymal stem cells either syngeneic to host or to donor, are effective on increasing survival time of graft; but in just a single dosage they are unable to induce specific immune tolerance.

Mahdokht Mahmoodi has completed her MSc in Medical Biotechnology from Shiraz University of Medical Sciences and Bachelor of Physiotherapy from Shahid Beheshti University of Medical Sciences, Tehran, Iran. She is working in Burn and Wound healing research center, Shiraz University of Medical Sciences as Research Assistant in Tissue Engineering and Cell Culturer laboratory form 2015 till present.

In recent years new approaches for wound healing are applied using new biotechnological and bioengineering methods which are very effective in regenerative medicine and tissue engineering. Grows factors play an important role in the healing process. Platelet Rich Plasma (PRP) is of the natural grows factors that are safe to use. Fibroblast cells are of the most effective cells in the wound healing process with the production of extracellular matrix and collagen to accelerate wound healing and also they are the most important cells in the proliferative phase. Studies have shown that microRNA can be used as a reducing agent or additive participate in the process of inflammation and regeneration of the epidermis, which is one of the most important stages of healing to participate speed up wound healing. In this study we treated fibroblast cells which were developed from human skin with PRP extracted from cord blood in different concentrations (1%, 5%, 10%, 15% and 20%). After 24h total RNA were extracted from treated fibroblast cells. Expression of miRNA21 and miRNA146a as effective miRNAs in fibroblast cells grows was detected using Real-time PCR after cDNA synthesis from extracted RNAs. Platelet Rich Plasma is a biological source of grows factors that affect fibroblast proliferation by enhancing miRNA21 and miRNA 146a as effective factors in wound healing. According to obtained results from this study fibroblast cells which were treated with 1%, 5% and 10% concentration of cord blood PRP showed expression of miRNA 21 in the 30-35th cycles of Real-time PCR and miRNA146a was expressed in a bit lower amount rather than miRNA21 in the 38-40th cycles of real-time PCR. As results showed, PRP stimulated expression of grows factors such as miRNA21 and miRNA 146a in fibroblasts and could be effective for enhance wound healing.

Developing effective stem cell-based therapies requires complex isolation of stem cells for in vitro culture systems, for accurate representation of the physiological stem cell niche. Even though bone marrow remains to be the gold standard for mesenchymal stem cells (MSCs) isolation, it is still considered a very invasive procedure. In an attempt to identify potential alternative MSCs sources for therapeutic applications, we studied MSCs derived from Wharton's Jelly of Umbilical Cord (WJUC) being an easy accessible source. We investigated their potency for in vitro expansion and differentiation into renal cells by comparing the usage of different cocktails as nephrogenic growth factors and basal medium as control. MSCs were isolated from 6 human UCs and further immunophenotyping using immunocytochemistry and CD29 expression flow cytometry were performed. In vitro expansion and differentiation of these isolated cells into renal progenitor stem cells was performed by adding separately 5 different nephrogenic growth factor cocktails and basal medium, then assessing and comparing the outcomes through investigating their differentiation capacities. The WJ-UCs isolated MSCs, showed significant positive expression of the MSCs surface marker, CD29. After expansion and induction of differentiation, results revealed that using nephrogenic growth factors cocktail containing a combination of Retinoic acid + Activin-A, then further adding of bone morphogenetic proteins (BMP-7), gave the highest significant positive expression of renal progenitor stem cell surface markers CD24 and CD133 using immunocytochemistry and flow cytometry. WJ-UC seems to be a promising source alternative to BM-MSCs, since they can be easily accessible, without any ethical concerns and are considered as a medical waste. After in vitro expansion and differentiation, using appropriate studied growth factors cocktail, newly formed renal progenitor stem cells may be further utilized by infusion or injection, opening new perspectives for cell-based therapies in patients suffering from renal diseases.

Globally, there is over a million new annual cases of prostate cancer. In approximately 70% of the cases, at least one copy of the tumor suppressor gene, phosphatase and tensin homolog deleted on chromosome 10 (PTEN), is found to be lost at diagnosis. PTEN is a phosphatase that works in contrast to the class I PI3Ks (phosphoinositide 3-kinase). Class I PI3Ks are heterodimers composed of a catalytic subunit (p110α, p110β, p110δ, or p110γ), which lend their names to the different PI3K complexes, and a regulatory subunit (p85α, p85β, p84, or p101). These heterodimers are expressed differently and seem to assume specific roles in different cellular functions, both within different tissues and single cell types, and are able convey spatially restricted signals, by phosphorylating the PI(4,5)P2 (phosphatidylinositol 4,5-bisphosphate) to produce PI(3,4,5)P3 (phosphatidylinositol 3,4,5-trisphosphate) in the inner leaflet of plasma membrane. These lipids belong to a complex signaling network widely implicated in human pathophysiology. PI(3,4,5)P3 activates vital downstream effector proteins, namely Akt, which is crucial for many cellular processes such as glucose metabolism, transcription, cell proliferation, cell migration, and apoptosis. The PI(3,4,5)P3 pool is controlled by the dephosphorylating abilities of PTEN, to produce PI(4,5)P2, and SHIP to produce PI(3,4)P2, respectively. PI(3,4)P2 is an important second messenger itself that can also activate Akt. The loss of PTEN causes unregulated PI3K/Akt signaling, which allow survival of prostate cancer cells and prevent apoptosis. We have utilized an established system to culture mouse prostate organoids from prostate progenitor cells to explore the role PI3K signaling in prostate growth, metabolism and tumorigenesis. The prostate-specific deletion of PTEN results in a significant reduction in the PTEN gene and protein expression in the mouse prostate tissue and cultured organoids. The animals display hyperplasic growth of epithelial cells, which develops into prostate intraepithelial neoplasia by 6-8 weeks, and eventually to adenocarcinoma by 4 months of age. Whilst, cultured organoids grow to a condensed yet sizable cell mass, in comparison to the wild type controls. This presented a good opportunity to study the PI3K signaling mechanisms of prostate cancer, and the tumor microenvironment in isolation. Our data indicates that there is a substantial increase in the accumulated levels of PI(3,4,5)P3 and PI(3,4)P2 lipids. This may offer a route in which these lipids play a role in the development and survival of prostatic cancerous cells. We show that this increased level of signaling is primarily driven by the PI3Kα isoform. We further report that the aberrant signaling is dependent on the pH level of the tumor microenvironment, as raising the pH of the organoids medium dramatically increases accumulation of PI(3,4,5)P3 and PI(3,4)P2, although the cause of this effect was unclear, we hypothesize the pH of the local environment may influence signaling via class I PI3Ks. Furthermore, the vast accumulation of PI(3,4)P2 -seen suggest that PTEN may also negatively regulate the PI(3,4)P2 signal, in addition to that of PI(3,4,5)P3. Our findings further our knowledge in understanding the mechanisms of this disease and contribute to the development of more targeted treatments.