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Introduction: Type 2 diabetes, caused by an insulin resistant, is associated with endothelial cell dysfunction and impaired vascular homeostasis, resulting in vascular inflammation and atherosclerosis. Recently, endothelial progenitor cells (EPCs) which originated from the bone marrow are considered to be an important regulator in vascular homeostasis and a surrogate marker for vascular complications. According to this, EPC dysfunction may play an important role in causing the diabetic-associated vascular complications. Objective: To compare number and characteristic of EPCs in Type 2 diabetic patients with those of normal healthy subjects. Methods: The number of EPCs from twenty Type 2 diabetic patients and twenty healthy subjects was studied by using flow cytometry. The isolated EPCs were cultured and characterized by Dil-AcLDL engulfment. The various glucose concentrations were employed in the EPC culture in order to determine the effect of hyperglycemia on the number and viability of the EPCs. Results: The number of EPCs in Type 2 diabetic patients was significantly decreased compared to healthy subjects (5.5 \× 10⁶ \± 0.5 \× 10⁶ vs. 23 \× 10⁶ \± 2.3 x 10⁶; P \< 0.01). There was an inverse correlation between the EPC numbers and the concentrations of plasma glucose (r = -0.045) as well as HbA1c (r = -0.336). The culture of EPCs from diabetic patients took a significantly longer period of time to develop mature EPCs than control (30.8 \± 3.9 days vs. 22.4 \± 2.7 days, P \< 0.01). In addition, the number of cultured EPCs was significantly reduced when the glucose concentration in culture medium was greater than 16.5 mmol/l (P \< 0.05). These results demonstrated that glucose has a negative effect on the number and viability of EPCs in a dose-dependent fashion. Conclusion Hyperglycemic condition in Type 2 diabetes has a negative effect on the number and viability of circulating EPCs in a dose-dependent fashion.
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The semen preparation medium that yields good semen quality is one among various important factors in the process of infertility treatment. The objective of this study was to compare the semen analysis parameters before and after preparation with 3 semen preparation media i.e. PureSperm® (Nidacon, Gothenburg, Sweden), Sil-Select Plus™ (Fertipro, Beernem, Belgium) and SpermGrad™ (Vitrolife, Gothenburg, Sweden) for enhancement of good semen quality.The total of 28 males of infertile couples attending Fertility Clinic of Thammasat University Hospital during the year 2007 were studied. Their semen (pre-washed) were analyzed for semen analysis parameters according to WHO¹ and Kruger’ strict criteria² and then equally divided into 3 aliquots and processed with 3 semen preparation media followed by semen analysis once again (post-washed). The sperm concentration, motility, progressive motile concentration (PMC), morphology, DNA damage and protamine deficiency of before and after preparation were compared by paired t-test with significance at P \< 0.05.The mean percentage of sperm motility, PMC, rapid motility and normal morphology were significantly increased where the mean percentage of slow motility, sperm head defect, sperm concentration and protamine deficiency were significantly decreased after preparation with all 3 media. The DNA damaged sperm was significantly decreased in the post-washed with PureSperm® and Sil-Select Plus™ but significantly increased with SpermGrad™.From the results obtained in this study, it could be concluded that PureSperm® may be suitable for both IUI and IVF techniques, since it yielded the best motile sperm and it may be also good for ICSI because it gave low percentage of DNA damaged sperm. Sil-Select Plus™ is probably best for ICSI since it gave lowest percentage of DNA damage sperm, whereas SpermGrad™ had highest percentage of DNA damage, thus, it is surely not suitable for ICSI procedure. Anyhow, SpermGrad™ may be moderately good for IUI since it had high percentage of motility and of normal morphology with lowest percentage of head defect but percentage of abnormal embryo may be increased.
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Objective: To isolate and characterize mesenchymal stem cells (MSCs) from peripheral blood and G-CSF mobilized peripheral blood. Methods: Mononuclear cells (MNCs) were isolated from peripheral blood, G-CSF mobilized peripheral blood and bone marrow using gradient centrifugation. The numbers of MSCs in these three sources were quantified using flow cytometry. The isolated MNCs were then cultured to generate MSCs. The MSCs generated from those three sources were studied in term of the MSC marker (CD73, CD90, CD105 and CD106) expression, the ability to generate colony (CFU-F) in culture and the ability to differentiate toward osteocyte and adipocyte-lineages. Results: The percentage of cells that expressed CD90 in fresh MNC populations isolated from bone marrow (BM-MNCs), peripheral blood (PB-MNCs) and mobilized peripheral blood (MPB-MNCs) is 1.94, 2.1 and 0.05 respectively, while the expression of CD73 in BM-MNCs, PB-MNCs and MPB-MNCs is 5.2, 15.2 and 7.8 respectively. The percentage of cells that expressed CD105 in BM-MNCs, PB-MNCs and MPB-MNCs is 5.3, 4.1 and 2.75, respectively while the expression of CD106 in those three populations is 2.82, 2.36 and 4.5 respectively. The ability of BM-MNCs, PB-MNCs and MPB-MNCs to generate colony in culture (CFU-F) is 67, 30 and 48 colonies per 10⁶ plating MNCs, respectively. After culture for three passages, more than 64% of BM-MNCs, PB-MNCs and MPB-MNCs homogeneously expressed CD73, CD90 and CD105. In contrast, the expression of CD45 (marker of hematopoietic cells) in those populations is negative. In addition, the bone marrow-derived MSCs also have an ability to differentiate toward osteocyte and adipocyte-lineages. Conclusion: We have successfully isolated and characterized MSCs from both peripheral blood and G-CSF mobilized peripheral blood. Those MSCs expressed several MSC markers, including CD73, CD90, CD105 and CD106, and able to generate colonies in culture in a manner similar to those of BM-MSCs. Our results suggest that these PB-MSCs and MPB-MSCs might be used as an alternative source for the clinical treatment in the future.
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Induced pluripotent stem (iPS) cells result from a reprogramming of somatic cells via force expression of the genes, Oct3/4, Sox2, c-myc and Klf4, which are essential for the establishment and maintenance of the pluripotent state. In properties, iPS cells are almost fully similar to embryonic stem (ES) cells. To date, iPS cells have been obtained from various differentiated cells of mice and human. In comparison with ES cells, iPS cells represent one of the most promising sources of patient-specific stem cells for use in research and regenerative medicine.
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Mesenchymal stem cells (MSCs) are capable of differentiating into different mesenchymal lineages, including adipose tissue, bone and cartilage. For clinical use, adult bone marrow is the most common source of MSCs. However, the frequency and differentiating capacity of MSCs in adult bone marrow decrease with age of donors. Different post-natal tissues have been studied for the presence of MSCs. However, there is no report about the characteristic of MSCs isolated from those sources in comparison to MSCs isolated from bone marrow. This study comprehensively compared several characteristics of MSCs isolated from umbilical cord, Wharton’s jelly and bone marrow in aspects of cell morphology, immunophenotype and growth kinetics as well as the differentiation capacity to be adipogenic and osteogenic lineages. We successfully isolated MSCs from umbilical cord and Wharton’s jelly, as well as bone marrow. Our results indicated that MSCs isolated from those three sources have fibroblast-like morphology and exhibited similar cell surface markers including CD73, CD90, and CD105. Umbilical cord and Wharton’s jelly derived MSCs cultured in NH OsteoDiff Medium and NH AdipoDiff Medium showed the ability to differentiate to be osteocytes and adipocytes in a comparable degree to bone marrow derived MSCs. Moreover, the proliferation capacity of MSCs isolated from umbilical cord and Wharton’s jelly at passage 2 to 4 was significantly higher than bone marrow derived MSCs (p \< 0.05). The results obtained from this study indicated that MSCs isolated from umbilical cord and Wharton’s jelly had similar characteristic in comparison to bone marrow derived MSCs. Therefore, post-natal tissues, especially umbilical cord and Wharton’s jelly, are attractive alternative sources of MSCs for future clinical application.