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Induction of Nestin Early Expression as a Hallmark for Mesenchymal Stem Cells Expression of PDX-1 as a Pre-disposing Factor for Their Conversion into Insulin Producing Cells.

Marisela MARTINEZ-GAMBOA ; Delia Elba CRUZ-VEGA ; Jorge MORENO-CUEVAS ; Maria Teresa GONZALEZ-GARZA

International Journal of Stem Cells.2017;10(1):76-82. doi:10.15283/ijsc16040

Diabetes constitutes a worldwide epidemic that affects all ethnic groups. Cell therapy is one of the best alternatives of treatment, by providing an effective way to regenerate insulin-producing cells lost during the course of the disease, but many issues remain to be solved. Several groups have been working in the development of a protocol capable of differentiating Mesenchymal Stem Cells (MSCs) into physiologically sound Insulin Producing Cells (IPCs). In order to obtain a simple, fast and direct method, we propose in this manuscript the induction of MSCs to express NESTIN in a short time period (2 h), proceeded by incubation in a low glucose induced medium (24 h) and lastly by incubation in a high glucose medium. Samples from cell cultures incubated in high glucose medium from 12 to 168 h were obtained to detect the expression of INSULIN-1, INSULIN -2, PDX-1 and GLUT-2 genes. Induced cells were exposed to a glucose challenge, in order to assess the production of insulin. This method allowed us to obtain cells expressing PDX-1, which resembles a progenitor insulin-producing cell.
Cell Culture Techniques ; Cell- and Tissue-Based Therapy ; Ethnic Groups ; Glucose ; Humans ; Insulin* ; Mesenchymal Stromal Cells* ; Methods ; Nestin*

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Bone Marrow Stem Cells Anti-liver Fibrosis Potency: Inhibition of Hepatic Stellate Cells Activity and Extracellular Matrix Deposition.

Ervina Julien SITANGGANG ; Radiana Dhewayani ANTARIANTO ; Sri Widia A JUSMAN ; Jeanne Adiwinata PAWITAN ; Ahmad Aulia JUSUF

International Journal of Stem Cells.2017;10(1):69-75. doi:10.15283/ijsc16048

Transplantation of bone marrow derived stem cells (BMSCs) has been reported inhibits liver fibrosis. Several in vitro studies by co-culturing BMSCs and hepatic stellate cells (HSCs) indirectly or directly in 2D models showed inhibition of HSC as the key player in liver fibrosis. In this study, we investigated direct effect of BMSCs on HSCs by co-culturing BMSCs and HSCs in 3D model as it represents the liver microenvironment with intricate cell-cell and cell-matrix interactions. Primary isolated rat HSCs and BMSCs were directly co-cultured at 1:1 ratio with hanging drop method. The monoculture of rat HSCs served as positive control. Mono-culture and co-culture samples were harvested on day 3, 5 and 7 for histological analysis. The samples were analyzed for extracellular matrix deposition by Masson's Trichrome staining, tenascin-C immunocytochemistry, resting HSC's state as shown by positive Oil Red O stained cells. Our results indicated CD90+CD34− BMSCs anti-liver fibrosis potency as evidenced by higher proportion of Oil Red O-positive cells in the co-culture group compared to the monoculture group and the significant decrease in extracellular matrix deposition as well as the decrease in tenascin-C expression in the co-culture group (p<0.05) compared to the monoculture group. These findings demonstrate that BMSCs have a potential therapeutic effect against liver fibrotic process through their capacity to inhibit HSCs activation and their effect in minimizing extracellular matrix deposition.
Animals ; Bone Marrow* ; Coculture Techniques ; Extracellular Matrix* ; Fibrosis* ; Hepatic Stellate Cells* ; Immunohistochemistry ; In Vitro Techniques ; Liver ; Liver Cirrhosis ; Methods ; Rats ; Stem Cells* ; Tenascin

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Conditioned Medium Derived from Salidroside-Pretreated Mesenchymal Stem Cell Culture Ameliorates Mouse Lipopolysaccharide-Induced Cerebral Neuroinflammation-Histological and Immunohistochemical Study.

Zeinab M EL MAADAWI

International Journal of Stem Cells.2017;10(1):60-68. doi:10.15283/ijsc16055

BACKGROUND AND OBJECTIVES: Neuroinflammation is involved in the pathogenesis of neurodegenerative disorders. Conditioned medium (CM) derived from bone marrow mesenchymal stem cells (MSCs) revealed substantial benefits due to its rich content of trophic factors. Salidroside (Sal), extracted from Rhodiola rosea, is known for its anti-inflammatory and neuroprotective effects. This study was designed to investigate the effect of Sal pretreated CM (CM-Sal) derived from bone marrow MSCs in lipopolysaccharide (LPS) induced neuroinflammation. MATERIAL AND METHODS: Fifty adult male mice were equally divided into 5 groups: Group I (Normal Control), Group II (LPS): single 0.8 mg/kg LPS intraperitoneally; Group III (LPS-DMEM), Group IV (LPS-CM) and Group V (LPS-CM-Sal): LPS was injected as group II followed, 24 hours later, by intranasal injection of 50 μl of filtered serum-free Dulbecco's Modified Eagle's medium (DMEM), CM or CM-Sal, respectively, twice daily for 4 days. Animals were sacrificed at day 6 and paraffin cerebral sections were subjected to Hematoxylin and Eosin staining and immunohistochemistry with caspase 3 (apoptosis), glial fibrillary acidic protein GFAP (astrocytes) and CD68 (active microglia) followed by quantitative morphometric study. RESULTS: Examination of LPS and LPS-DMEM groups revealed neuronal apoptosis with reactive astrogliosis and increased active microglia. LPS-CM and LPS-CM-Sal groups showed less apoptosis, less astrocytes and less active microglia. The regression in neuroinflammation was more evident in LPS-CM-Sal group and the difference was statistically significant compared to other groups. CONCLUSION: CM-Sal derived from MSCs culture elicited significant histopathological improvement in LPS induced neuroinflammation which could be used as new therapeutic modality.
Adult ; Animals ; Apoptosis ; Astrocytes ; Bone Marrow ; Caspase 3 ; Culture Media, Conditioned* ; Eosine Yellowish-(YS) ; Glial Fibrillary Acidic Protein ; Hematoxylin ; Humans ; Immunohistochemistry ; Male ; Mesenchymal Stromal Cells* ; Mice* ; Microglia ; Neurodegenerative Diseases ; Neurons ; Neuroprotective Agents ; Paraffin ; Rhodiola

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Cardioprotective Effects of Wharton Jelly Derived Mesenchymal Stem Cell Transplantation in a Rodent Model of Myocardial Injury.

Taghrid GAAFAR ; Wael ATTIA ; Shereen MAHMOUD ; Dina SABRY ; Osama Abdel AZIZ ; Dina RASHEED ; Hala HAMZA

International Journal of Stem Cells.2017;10(1):48-59. doi:10.15283/ijsc16063

BACKGROUND: Whartons jelly-derived mesenchymal stem cells are a valuable alternative source that possess multipotent properties, easy to obtain and available in large scale compared to BMMSCs. We investigated the possibility of cardiac function improvement post isoproterenol induced cardiac injury in a rat model following human WJMSCs transplantation. MATERIALS AND METHODS: MSCs were extracted and cultured from cord WJ, characterized by morphology, Immunophenotyping and differentiation to osteoblast and adipocytes. WJMSCs were labeled with PKH2 linker dye. Wistar rats were divided into control group, ISO group (injected with 2 doses of isoproterenol) to induce myocardial injury and ISO group transplanted with labelled WJMSCs. ECG, electrocardiographic patterns, cardiac marker enzymes, tracing of labeled MSCs and immunohistochemical analysis of myocardial cryosections were studied. RESULTS AND CONCLUSIONS: WJ derived MSCs were expanded for more than 14 passages while maintaining their un-differentiated state, were positive for MSC markers and were able to differentiate into adipocyte and osteoblast. We demonstrated that intravenously administered WJMSCs were capable of homing predominently in the ischemic myocardium. Cardiac markers were positively altered in stem cell treated group compared to ISO group. ECG and ECHO changes were improved with higher survival rate. WJMSCs could differentiate into cardiac-like cells (positive for cardiac specific proteins) in vivo. WJMSCs infusion promoted cardiac protection and reduced mortality, emphasizing a promising therapeutic role for myocardial insufficiency.
Adipocytes ; Electrocardiography ; Humans ; Immunophenotyping ; Isoproterenol ; Mesenchymal Stem Cell Transplantation* ; Mesenchymal Stromal Cells* ; Models, Animal ; Mortality ; Myocardium ; Osteoblasts ; Rats, Wistar ; Rodentia* ; Stem Cells ; Survival Rate ; Transplantation ; Wharton Jelly*

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Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells.

Mahmoud A SAEED ; Mohamed Abd EL-RAHMAN ; Mohamed E HELAL ; Ahmed R ZAHER ; Mohammed E GRAWISH

International Journal of Stem Cells.2017;10(1):38-47. doi:10.15283/ijsc16067

BACKGROUND AND OBJECTIVES: The imperative role of dental pulp stem cells (DPSCs) in regenerative therapy demands an in-vitro expansion which must deal with the safety and ethical problems associated with fetal bovine serum (FBS). The primary aim of this study was to compare the effects of human platelet rich fibrin (hPRF) exudate Vs FBS on proliferation and osteodifferentiation of human dental pulp stem cells (hDPSCs). The secondary one was to determine the optimum concentration of hPRF exudate inducing hDPSCs proliferation and osteodifferentiation. METHODS: The direct method was used to prepare hPRF exudate. hDPSCs were isolated from impacted mandibular third molars of twelve donors by the outgrowth method. For cell viability and proliferation rate testing, 96 well plates were used and the assay was done in duplicate and the trial repeated four times under the same conditions. Six wells were used to contain 10% FBS, serum free media, 1%, 5%, 10% and 20% concentrations of hPRF exudates, respectively. The proliferation assay was carried out by MTS tetrazolium cell proliferation assay kit and Elisa reader. The study design for osteodifferentiation protocol was exactly as the proliferation one and instead the assay was carried out by alizarin red with Elisa reader. RESULTS: Compared to 10% FBS, 10% hPRF exudate was the optimum concentration for hDPSCs proliferation, while 1% hPRF exudate was the optimum concentration for osteodifferentiation of hDPSCs. CONCLUSIONS: Avoiding the risk of zoonosis which may be occurred with FBS, it is recommended to use 10% hPRF exudate for proliferation and 1% for osteodifferentiation.
Blood Platelets* ; Cell Proliferation ; Cell Survival ; Culture Media, Serum-Free ; Dental Pulp* ; Enzyme-Linked Immunosorbent Assay ; Exudates and Transudates* ; Fibrin* ; Humans* ; Methods ; Molar, Third ; Stem Cells* ; Tissue Donors

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Lineage-specific Expression of miR-200 Family in Human Embryonic Stem Cells during In Vitro Differentiation.

Yeji KIM ; Nury KIM ; Sang Wook PARK ; Hyemin KIM ; Han Jin PARK ; Yong Mahn HAN

International Journal of Stem Cells.2017;10(1):28-37. doi:10.15283/ijsc17013

Although microRNAs have emerged as key regulators in diverse cellular processes, the roles of microRNAs are poorly understood in human embryonic stem cells (hESCs) during differentiation into specialized cell types. In this study, we used a microRNA array with 799 human microRNA probes to examine the expression profiles of microRNAs in hESCs during differentiation into endodermal and mesodermal lineages in vitro. Among the microRNAs analyzed, 7 and 20 microRNAs were enriched in the developmental process of hESCs into mesodermal and endodermal lineages, respectively. In particular, the expression levels of miR-200 family, which is known to regulate the epithelial to mesenchymal transition (EMT), gradually increased in hESCs during differentiation into hepatocytes while they gradually decreased during differentiation into vascular endothelial cells. Downregulation of ZEB1, a direct target of miR-200 family, and E-CADHERIN, a target protein of ZEB1, was observed in hESCs during differentiation into endodermal and mesodermal lineages, respectively. These results indicate that miR-200 family has an important role in determining the cell fate between endodermal and mesodermal lineages from the pluripotent state.
Cadherins ; Down-Regulation ; Endoderm ; Endothelial Cells ; Hepatocytes ; Human Embryonic Stem Cells* ; Humans ; Humans* ; In Vitro Techniques* ; Mesoderm ; MicroRNAs

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Wasim KAMATE ; Rajendra BAAD ; Nupura VIBHUTE ; Uzma BELGAUMI ; Vidya KADASHETTI ; Sushma GUGWAD

International Journal of Stem Cells.2017;10(1):21-27. doi:10.15283/ijsc16065

Tumor-initiating cells are a diminutive subpopulation of stem cells that have ability of long term self-renewal and generation of varied traits of tumor cell population. Understanding the concept of tumor-initiating cells may have a great implicative intimation for our comprehension of cancer pathobiology and for the delineation of new therapies directed towards these stem cells. The present review is an endeavor to conceptualize the role of tumor-initiating cells in the Squamous Cell Cancers (SCC) of head and neck, their role in tumorigenesis and the possible supplementary approach in the latest treatment modalities.
Carcinogenesis ; Comprehension ; Epithelial Cells* ; Head* ; Neck* ; Neoplasms, Squamous Cell* ; Stem Cells

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Methods of Isolation and Characterization of Stem Cells from Different Regions of Oral Cavity Using Markers: A Systematic Review.

Kavarthapu AVINASH ; Sankari MALAIPPAN ; Jayakumar Nadathur DOORAISWAMY

International Journal of Stem Cells.2017;10(1):12-20. doi:10.15283/ijsc17010

BACKGROUND: Periodontitis is a destructive inflammatory disorder of the periodontium caused by the destruction of periodontal tissues namely the PDL, cementum, alveolar bone, and gingiva. Once these tissues are lost, the foremost goal of periodontal therapy is to regenerate the diseased tissues if possible to their original form, architecture, and function. Various regenerative procedures were employed and still a gap was found in achieving the goal. As stem cells are characterized by their ability to self-renew and differentiate to produce specialized cells, there could be a possibility of using them for regenerative therapy. Recently, dental tissues such as the PDL, the dental pulp and the tooth follicle have been recognized as readily available sources of adult stem cells. AIM: The aim was to identify the various sources and methodologies in isolation of stem cells from human oral cavity and its differentiation into various lineages using markers. MATERIALS AND METHODS: The electronic databases PUBMED, GOOGLE SCHOLAR, SCIENCE DIRECT, COCHRANE LIBRARY along with a complimentary manual search of all periodontics journal till the year 2016. Thirteen articles were selected on the basis of the inclusion criteria. Isolation of stem cells from oral cavity through various methods has been evaluated and similarly characterization to different lineages were tabulated as variables of interest. They included human in-vitro and ex-vivo studies. RESULTS: The results showed that PDLSC's and pulpal stem cells are the most common source from where stem cells were isolated. Each source has used different methodology in isolating the stem cells and it was found that STRO-1 was the commonly used marker in all the studies mentioned. CONCLUSIONS: The studies showed that there is no standard protocol existed in isolating the stem cells from different sources of oral cavity. Moreover, there was no standard marker or methodology used in characterization.
Adult Stem Cells ; Dental Cementum ; Dental Pulp ; Gingiva ; Humans ; Methods* ; Mouth* ; Periodontics ; Periodontitis ; Periodontium ; Stem Cells* ; Tooth

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From Bench to Market: Preparing Human Pluripotent Stem Cells Derived Cardiomyocytes for Various Applications.

Sung Hwan MOON ; Daekyeong BAE ; Taek Hee JUNG ; Eun Bin CHUNG ; Young Hoon JEONG ; Soon Jung PARK ; Hyung Min CHUNG

International Journal of Stem Cells.2017;10(1):1-11. doi:10.15283/ijsc17024

Human cardiomyocytes (CMs) cease to proliferate and remain terminally differentiated thereafter, when humans reach the mid-20s. Thus, any damages sustained by myocardium tissue are irreversible, and they require medical interventions to regain functionality. To date, new surgical procedures and drugs have been developed, albeit with limited success, to treat various heart diseases including myocardial infarction. Hence, there is a pressing need to develop more effective treatment methods to address the increasing mortality rate of the heart diseases. Functional CMs are not only an important in vitro cellular tool to model various types of heart diseases for drug development, but they are also a promising therapeutic agent for cell therapy. However, the limited proliferative capacity entails difficulties in acquiring functional CMs in the scale that is required for pathological studies and cell therapy development. Stem cells, human pluripotent stem cells (hPSCs) in particular, have been considered as an unlimited cellular source for providing functional CMs for various applications. Notable progress has already been made: the first clinical trials of hPSCs derived CMs (hPSC-CMs) for treating myocardial infarction was approved in 2015, and their potential use in disease modeling and drug discovery is being fully explored. This concise review gives an account of current development of differentiation, purification and maturation techniques for hPSC-CMs, and their application in cell therapy development and pharmaceutical industries will be discussed with the latest experimental evidence.
Cell- and Tissue-Based Therapy ; Drug Discovery ; Drug Industry ; Heart Diseases ; Humans* ; In Vitro Techniques ; Mortality ; Myocardial Infarction ; Myocardium ; Myocytes, Cardiac* ; Pluripotent Stem Cells* ; Stem Cells

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FGF8 is Essential for Functionality of Induced Neural Precursor Cell-derived Dopaminergic Neurons.

Mi Sun LIM ; Soo Young LEE ; Chang Hwan PARK

International Journal of Stem Cells.2015;8(2):228-234. doi:10.15283/ijsc.2015.8.2.228

Induced neural precursor cells (iNPCs) are one source of transplantable dopaminergic neurons used in cell therapy for Parkinson's disease. In the present study, we demonstrate that iNPCs can be generated by transducing Brn2, Ascl1, Myt1L and Bcl-xL in a culture supplemented with several mitogens and subsequently can be differentiated to dopaminergic neurons (DA). However, studies have shown that iDA and/or iNPC-derived DA neurons using various conversion protocols have low efficiency. Here, we show that early exposure of FGF8 to fibroblasts efficiently improves differentiation of DA neurons. So our study demonstrates that FGF8 is a critical factor for generation of iNPC-derived DA neurons.
Cell- and Tissue-Based Therapy ; Dopaminergic Neurons* ; Fibroblasts ; Mitogens ; Neurons ; Parkinson Disease

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