No abstract available.
Embryonic Stem Cells*

BACKGROUND: This study was designed to verify the effective culture condition for the differentiation of human hematopoietic stem cells from SNU-3 embryonic stem cell line. METHODS: Control group was that of which embryonic stem cells were directly cultured to LTC-IC assay. Study group was that of which an embryonic body manufactured from embryonic stem cells was cultured to LTC-IC assay. CD34+ cells were separated by MACS method at 1, 3, 5, 7, 10, 14, 21 days of LTC-IC assay in both groups. Thereafter, CD34+ cell were cultured to semisolid methyl-cellulose media to count CFUs. CD34+CD45- cell percentage was measured with FACS method at 5 days of LTC-IC assay. This study was repeated for 14 times. RESULTS: In control group, CD34+ cells were hardly separated in any period of LTC-IC assay. In study group, the median count of CD34+ cells was 0.7 (0.4-1.2)x10(4), 1.8 (1.4-2.6)x10(4), 0.6 (0.5-0.7)x10(4) and the median count of CFUs was 0.5 (0.2-0.8)x10(2), 1.0 (0.6-1.3)x10(2), 0.2(0.1-0.4)x10(2) at 3, 5, 7 days of LTC-IC assay, respectively. Median CFUs count per CD34+ cell was 0.0071, 0.0056, 0.0033 at 3, 5, 7 days of LTC-IC assay, respectively. In study group, the count of CD34+ cells and CFUs was significantly higher at 5 days of LTC-IC assay than at any other days (P<0.01). CFUs count per CD34+ cell was significantly higher at 3 days of LTC-IC assay than at any other days (P<0.01). CD34+CD45- cells detected by FACS method of study group(1.16%(0.92-1.97) was significantly higher than that of control group (0.09% (0.00-0.23)) (P<0.01). CONCLUSION: The differentiation of hematopoietic stem cells from SNU-3 embryonic stem cell line is effective in the condition of which an embryonic body manufactured from embryonic stem cells is cultured to LTC-IC assay. The period of which embryonic stem cells differentiate to hematopoietic stem cells is between 3 to 7 days of LTC-IC assay.
Embryonic Stem Cells* ; Hematopoietic Stem Cells* ; Humans

No abstract available.
Embryonic Stem Cells ; Neural Crest

Embryonic stem cells can be differentiated into various types of cells, requiring a tight regulation of transcription. Biomarkers related to each lineage of cells are used to guide the differentiation into neural or any other fates. In previous experiments, we reported the guided differentiation (GD)-specific genes by comparing profiles of random differentiation (RD). Interestingly 68% of differentially expressed genes in GD overlap with that of RD, which makes it difficult for us to separate the lineages by examining several markers. In this paper, we design a prediction model to identify the differentiation into neural fates from any other lineage. From the profiles of 11,376 genes, 203 differentially expressed genes between neural and random differentiation were selected by random variance T-test with 95% confidence and 5% false discovery rate. Based on support vector machine algorithm, we could select 79 marker genes from the 203 informative genes to construct the optimal prediction model. Here we propose a prediction model for the prediction of neural fates from random differentiation which is constructed with a perfect accuracy.
Embryonic Stem Cells ; Stem Cells ; Support Vector Machine ; Biomarkers

Induced pluripotent stem cells (iPSCs) are a type of cells similar to embryonic stem cells but produced by reprogramed somatic cells. Through in vitro differentiation of iPSCs, we can interrogate the evolution history as well as the various characteristics of macrophages. iPSCs derived macrophages are not only a good model for drug screening, but also an important approach for immunotherapy. This review summarizes the advances, challenges, and future directions in the field of iPSCs-derived macrophages.
Cell Differentiation ; Embryonic Stem Cells ; Induced Pluripotent Stem Cells ; Macrophages

No abstract available.
Cryopreservation* ; Embryonic Stem Cells* ; Humans* ; Vitrification

Human Embryonic Stem Cells ; Humans ; Safety

Human embryonic stem cell (hESC) culture system has been changing culture conditions from conventional to xeno-free for therapeutic cell applications, and N-glycolylneuraminic acid (Neu5Gc) could be a useful indicator of xenogeneic contaminations in hESCs because human cells can no longer produce it genetically. We set up the humanized culture condition using commercially available humanized materials and two different adaptation methods: sequential or direct. SNUhES4 and H1 hESC lines, previously established in conventional culture conditions, were maintained using the humanized culture condition and were examined for the presence of Neu5Gc. The hESCs showed the same morphology and character as those of the conventional culture condition. Moreover, they were negative for Neu5Gc within two passages without loss of pluripotency. This study suggested that this method can effectively cleanse previously established hESC lines, bringing them one step closer to being clinical-grade hESCs.
Human Embryonic Stem Cells* ; Humans* ; Methods

The study aims to investigate the effects of cardiac fibroblast (CF) paracrine factors on murine embryonic stem cells (ESCs). Conditioned mediums from either neonatal cardiac fibroblasts (ConM-NCF) or adult cardiac fibroblasts (ConM-ACF) were diluted by 1:50 and 1:5, respectively, to investigate whether these conditioned mediums impact murine ESCs distinctly with RT-real time PCR techniques, cell proliferation essay, ELISA and by counting percentage of beating embryoid bodies (EBs) during ESCs differentiation. The data showed that the paracrine ability of CFs changed dramatically during development, in which interleukin 6 (IL6) increased with maturation. ConM-NCF 1:50 and ConM-NCF 1:5 had opposite effects on the pluripotent markers, although they both reduced mouse ESC proliferation. ConM-ACF 1:50 promoted ESCs pluripotent markers and proliferation, while ConM-ACF 1:5 exerted negative effects. All CF-derived conditioned mediums inhibited cardiac differentiation, but with distinguishable features: ConM-NCF 1:50 slightly decreased the early cardiac differentiation without altering the maturation tendency or cardiac specific markers in EBs at differentiation of day 17; ConM-ACF 1:50 had more significant inhibitory effects on early cardiac differentiation than ConM-NCF 1:50 and impeded cardiac maturation with upregulation of cardiac specific markers. In addition, IL6 neutralization antibody attenuated positive effect of ConM-ACF 1:50 on ESCs proliferation, but had no effects on ConM-NCF 1:50. Long-term IL6 neutralization reduced the percentage of beating EBs at early developmental stage, but did not alter the late cardiac differentiation. Taken together, both the quality and quantity of factors and cytokines secreted by CFs are critical for the ESC fate. IL6 could be a favorable cytokine for ESC pluripotency and the early cardiac differentiation.
Animals ; Embryonic Stem Cells ; Fibroblasts ; Heart ; Mice ; Mouse Embryonic Stem Cells ; Paracrine Communication

Embryonic Stem Cells ; Humans ; Myocardial Infarction ; therapy ; Stem Cell Transplantation