Djuwantono T.,Padjadjaran University |
Wirakusumah F.F.,Padjadjaran University |
Achmad T.H.,Padjadjaran University |
Sandra F.,Stem Cell and Cancer Institute |
And 3 more authors.
BMC Research Notes | Year: 2011
Background: The finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Due to the once-in-a-life time chance of collecting it, an optimum cryopreservation method that can preserve the life and function of the cells contained is critically needed. Methods. Until now, slow-cooling has been the routine method of cryopreservation; however, rapid-cooling offers a simple, efficient, and harmless method for preserving the life and function of the desired cells. Therefore, this study was conducted to compare the effectiveness of slow- and rapid-cooling to preserve umbilical cord blood of mononucleated cells suspected of containing hematopoietic stem cells. The parameters used in this study were differences in cell viability, malondialdehyde content, and apoptosis level. The identification of hematopoietic stem cells themselves was carried out by enumerating CD34+ in a flow cytometer. Results: Our results showed that mononucleated cell viability after rapid-cooling (91.9%) was significantly higher than that after slow-cooling (75.5%), with a p value = 0.003. Interestingly, the malondialdehyde level in the mononucleated cell population after rapid-cooling (56.45μM) was also significantly higher than that after slow-cooling (33.25μM), with a p value < 0.001. The apoptosis level in rapid-cooling population (5.18%) was not significantly different from that of the mononucleated cell population that underwent slow-cooling (3.81%), with a p value = 0.138. However, CD34+ enumeration was much higher in the population that underwent slow-cooling (23.32 cell/μl) than in the one that underwent rapid-cooling (2.47 cell/μl), with a p value = 0.001. Conclusions: Rapid-cooling is a potential cryopreservation method to be used to preserve the umbilical cord blood of mononucleated cells, although further optimization of the number of CD34+ cells after rapid-cooling is critically needed. © 2011 Djuwantono et al; licensee BioMed Central Ltd.
PubMed | Stem cell and Cancer Institute, Bandung Institute of Technology, Hong Kong University of Science and Technology, Kalbe Genomics Laboratory and University of Melbourne
Type: | Journal: Journal of gastrointestinal cancer | Year: 2017
K-RAS and recently N-RAS gene mutation testing are mandatory requirements prior to anti-epidermal growth factor receptor (EGFR) monoclonal antibody treatment of metastatic CRC. Mutation prevalence and distribution in Indonesian colorectal cancer (CRC) are not known.Combined methods of PCR high-resolution melt (HRM), restriction fragment length polymorphism (RFLP), and direct DNA sequencing were used to genotype exons 2, 3, and 4 of both K-RAS and N-RAS genes for routine clinical testing of CRC patients. Descriptive analytical review of 595 consecutive CRC patients (years 2013 to 2016) was performed to find associations between gene mutations and clinicopathologic features.This retrospective study revealed overall K-RAS gene mutation in exon 2 (codon 12 and 13) rates being 34.9%. Women (42.5%), stages I and II (43.4%), and well and moderate differentiations (37.7%) had higher frequency of K-RAS exon 2 mutations than men (29%, p=0.006), stages (III and IV 31.9%, p=0.05), and poor differentiation (11.8%, p=0.002), respectively. At later period (2015-2016), 121 of 595 patients were genotyped for the remaining exons 3 and 4 of K-RAS as well as exons 2, 3, and 4 of N-RAS mutations resulting in overall RAS mutation prevalence of 41%. Mucinous histology had highest frequency of N-RAS mutation.Combination of PCR HRM with either RFLP or direct DNA sequencing was useful to detect K-RAS exon 2 and extended RAS mutations, respectively. Frequency of all RAS mutations in stage IV Indonesian (41%) was similar among Asians (41-49%), which tend to be lower than western (55%) CRC.
PubMed | Maranatha Christian University, Lambung Mangkurat University, Biomolecular and Biomedical Research Center, Pancasila University and 2 more.
Type: Journal Article | Journal: Iranian journal of basic medical sciences | Year: 2015
Many studies have reported that tea consumption decreases cardiovascular risk, but the mechanisms remain unclear. Green tea is known to have potent antioxidant and free radical scavenging activities. This study aimed to investigate whether green tea extract (GTE) can protect endothelial progenitors cells (EPCs) against oxidative stress through antioxidant mechanisms.Mononuclear cells (MNCs) were isolated from peripheral blood by density gradient centrifugation with Ficoll. The cells were then plated on fibronectin-coated culture dishes. After 7 days of culture, EPCs were characterized as adherent cells double positive for DiI-ac-LDL uptake and lectin binding. EPCs were further identified by assessing the expression of CD34/45, CD133, and KDR. EPCs were then treated with hydrogen peroxide (H2O2) at doses of 50, 100, 200 M and incubated with or without GTE (25 g/ml). The intracellular reactive oxygen species (ROS) levels were detected by flow cytometry using a 2,7-dichlorofluorescein diacetate (DCF-DA) fluorescent probe.GTE ameliorated the cell viability of EPCs induced by H2O2 at doses of 50, 100, 200 M for about 25.47, 22.52, and 11.96% higher than controls, respectively. GTE also decreased the intracellular ROS levels of EPCs induced by H2O2 at doses of 50, 100, 200 M for about 84.24, 92.27, and 93.72% compared to controls, respectively.GTE improves cell viability by reducing the intracellular ROS accumulation in H2O2-induced EPCs.
Widowati W.,Maranatha Christian University |
Wijaya L.,Stem Cell and Cancer Institute |
Bachtiar I.,Stem Cell and Cancer Institute |
Gunanegara R.F.,Maranatha Christian University |
And 4 more authors.
Biomarkers and Genomic Medicine | Year: 2014
Mesenchymal stem cells (MSCs) from Wharton's jelly have a higher proliferation rate and self-renewal capacity than adult tissue-derived MSCs. A low oxygen level or hypoxic condition is prevalent in the microenvironment of the stem cells in the early stages of development. Hypoxia can influence proliferation and differentiation of various stem/precursor cell populations. This research was conducted: to determine the proliferation rate and characteristics of human MSCs from Wharton's jelly in hypoxic and normoxic condition; to evaluate their character after MSCs are incubated in hypoxic and normoxic environment using surface markers including CD105, CD73, CD14, CD19, CD34, CD45, and HLA-II; and to evaluate the proliferation rate and number of MSCs at many passages using the trypan blue method. The hypoxic and normoxic microenvironment showed significant differences in the proliferation rate and population doubling time, but and there were no differences in surface markers. © 2014.
Li X.,Brigham and Women's Hospital |
Utomo A.,Brigham and Women's Hospital |
Utomo A.,Stem Cell and Cancer Institute |
Cullere X.,Brigham and Women's Hospital |
And 5 more authors.
Cell Host and Microbe | Year: 2011
Resistance to fungal infections is attributed to engagement of host pattern-recognition receptors, notably the β-glucan receptor Dectin-1 and the integrin Mac-1, which induce phagocytosis and antifungal immunity. However, the mechanisms by which these receptors coordinate fungal clearance are unknown. We show that upon ligand binding, Dectin-1 activates Mac-1 to also recognize fungal components, and this stepwise process is critical for neutrophil cytotoxic responses. Both Mac-1 activation and Dectin-1- and Mac-1-induced neutrophil effector functions require Vav1 and Vav3, exchange factors for RhoGTPases. Mac-1- or Vav1,3-deficient mice have increased susceptibility to systemic candidiasis that is not due to impaired neutrophil recruitment but defective intracellular killing of C. albicans yeast forms, and Mac-1 or Vav1,3 reconstitution in hematopoietic cells restores resistance. Our results demonstrate that antifungal immunity depends on Dectin-1-induced activation of Mac-1 functions that is coordinated by Vav proteins, a pathway that may localize cytotoxic responses of circulating neutrophils to infected tissues. © 2011 Elsevier Inc.
Wijaya L.,Stem Cell and Cancer Institute |
Agustina D.,Stem Cell and Cancer Institute |
Lizandi A.O.,Stem Cell and Cancer Institute |
Kartawinata M.M.,Stem Cell and Cancer Institute |
Sandra F.,Stem Cell and Cancer Institute
Current Pharmaceutical Biotechnology | Year: 2011
Stem cells have an important role in cell biology, allowing tissues to be renewed by freshly created cells throughout their lifetime. The specific micro-environment of stem cells is called stem cell niche; this environment influences the development of stem cells from quiescence through stages of differentiation. Recent advance researches have improved the understanding of the cellular and molecular components of the micro-environment - or niche - that regulates stem cells. We point out an important trend to the study of niche activity in breast cancers. Breast cancer has long been known to conserve a heterogeneous population of cells. While the majority of cells that make up tumors are destined to differentiate and eventually stop dividing, only minority populations of cells, termed cancer stem cell, possess extensive self renewal capability. These cancer stem cells possess characteristics of both stem cells and cancer cells. Breast cancer stem cells reversal to breast somatic stem cells offer a new therapy, that not only can stop the spread of breast cancer cells, but also can differentiate breast cancer stem cells into normal breast somatic stem cells. These can replace damaged breast tissue. Nevertheless, the complexity of realizing this therapy approach needs further research. © 2011 Bentham Science Publishers Ltd.
Arung E.T.,Stem Cell and Cancer Institute |
Arung E.T.,Mulawarman University |
Wicaksono B.D.,Stem Cell and Cancer Institute |
Handoko Y.A.,Stem Cell and Cancer Institute |
And 4 more authors.
Journal of Natural Medicines | Year: 2010
In our screening projects for anticancer agents from natural resources, artocarpin [6-(3-methyl-1-butenyl)-5,2′,4′-trihydroxy-3-isoprenyl-7- methoxyflavone] isolated from wood of jack fruit (Artocarpus heterophyllus) showed potent cytotoxic activity on human T47D breast cancer cells. The mode of action of artocarpin was evaluated by its effect on cell viability, nuclear morphology, cell cycle progression, expression of protein markers for apoptosis, and mitochondrial membrane potential (Δψm). These results showed that artocarpin caused a reduction of cell viability in a concentration-dependent manner and an alteration of cell and nuclear morphology. Moreover, the percentage of the sub-G1 phase formation was elevated dose-dependently. Artocarpin induced activation of caspase 8 and 10 as indicated by stronger signal intensity of cleaved-caspase 8 and weaker signal intensity of caspase 10 markers detected after artocarpin treatment. In addition, we also noticed the activation of caspase 3 by artocarpin. There were negligible changes in mitochondrial membrane potential (Δψm) due to artocarpin treatment. All together, these data indicated that artocarpin induced apoptosis in T47D cells possibly via an extrinsic pathway. © 2010 The Japanese Society of Pharmacognosy and Springer.
Mahanani E.S.,Gadjah Mada University |
Mahanani E.S.,University of Indonesia |
Bachtiar I.,Stem Cell and Cancer Institute |
Ana I.D.,Gadjah Mada University
Key Engineering Materials | Year: 2016
For bone tissue engineering, corals have long history to be used as scaffold to promote bone regeneration. However the use of a lot of corals may damage their habitates. For this reason, a strategy to mimic coral in a synthetic form is needed. As an ideal scaffold, synthetic coral must provide structure and initial support for cell attachment and proliferation. The aim of this study was to investigate the attachment and proliferation of human Mesenchymal Stem Cells (h-MSC) on synthetic coral scaffold, to provide information on the behavior of h-MSC on the designated scaffold. Synthetic coral scaffolds were prepared from bovine gelatine and CaCO3 with 5:5 in 10% w/v concentration in aquadest. Sodium citrate was used as dispersant in the suspension. Gelatin-CaCO3 suspension was moulded in a plastic cover of 24 well plate, then freezed at-20°C for 24 hours, freeze dried for 24 hours and continued by dehydrothermal crosslinking for 72 hours. After the fabrication, synthetic coral scaffolds were subjects to cover the bottom of the well for cell culture. Human Mesenchymal Stem Cells (h-MSC) were seeded and divided into 2 groups, control group without scaffold and the one with scaffold. All groups were incubated for 3, 6, and 24 hours. Cells attatchment were determined by deduction of the cells unattached from total cells seeding. Proliferation of h-MSC were done in 3 groups ie., control group without scaffold, scaffold only and scaffold incorporated Platelet Rich Plasma (PRP) in the bottom of well. All groups were incubated for 24, 48 and 72 hours. Human Mesenchymal Stem Cells attached faster to synthetic coral scaffold than the control. Its proliferation behavior was faster in the scaffold incorporated PRP, showing better interaction of scaffold and cells with the incorporation of morphogenetic factor. © 2016 Trans Tech Publications, Switzerland.
PubMed | Stem Cell and Cancer Institute
Type: Journal Article | Journal: Current pharmaceutical biotechnology | Year: 2011
Stem cells have an important role in cell biology, allowing tissues to be renewed by freshly created cells throughout their lifetime. The specific micro-environment of stem cells is called stem cell niche; this environment influences the development of stem cells from quiescence through stages of differentiation. Recent advance researches have improved the understanding of the cellular and molecular components of the micro-environment--or niche--that regulates stem cells. We point out an important trend to the study of niche activity in breast cancers. Breast cancer has long been known to conserve a heterogeneous population of cells. While the majority of cells that make up tumors are destined to differentiate and eventually stop dividing, only minority populations of cells, termed cancer stem cell, possess extensive self renewal capability. These cancer stem cells possess characteristics of both stem cells and cancer cells. Breast cancer stem cells reversal to breast somatic stem cells offer a new therapy, that not only can stop the spread of breast cancer cells, but also can differentiate breast cancer stem cells into normal breast somatic stem cells. These can replace damaged breast tissue. Nevertheless, the complexity of realizing this therapy approach needs further research.
PubMed | Stem Cell and Cancer Institute and National University of Indonesia
Type: | Journal: Cardiology research and practice | Year: 2016
Background. Proangiogenic Hematopoietic Cells (PHC) which comprise diverse mixture of cell types are able to secrete proangiogenic factors and interesting candidate for cell therapy. The aim of this study was to seek for benefit in implantation of PHC on functional improvement in end stage coronary artery disease patients with advanced heart failure. Methods. Patients with symptomatic heart failure despite guideline directed medical therapy and LVEF less than 35% were included. Peripheral blood mononuclear cells were isolated, cultivated for 5 days, and then harvested. Flow cytometry and cell surface markers were used to characterize PHC. The PHC were delivered retrogradely via sinus coronarius. Echocardiography, myocardial perfusion, and clinical and functional data were analyzed up to 1-year observation. Results. Of 30 patients (56.4 7.40yo) preimplant NT proBNP level is 5124.5 4682.50pmol/L. Harvested cells characterized with CD133, CD34, CD45, and KDR showed 0.87 0.41, 0.63 0.66, 99.00 2.60, and 3.22 3.79%, respectively. LVEF was improved (22 5.68 versus 26.8 7.93, p < 0.001) during short and long term observation. Myocardial perfusion significantly improved 6 months after treatment. NYHA Class and six-minute walk test are improved during short term and long term follow-up. Conclusion. Expanded peripheral blood PHC implantation using retrograde delivery approach improved LV systolic function, myocardial perfusion, and functional capacity.