AIM： To investigate the expression of coxsackievirus and adenovirus receptor （CAR） and adenovirus-mediated reporter gene transfer in five human colon cancer cell lines.METHODS： Expression of CAR-specific mRNA and ...AIM： To investigate the expression of coxsackievirus and adenovirus receptor （CAR） and adenovirus-mediated reporter gene transfer in five human colon cancer cell lines.METHODS： Expression of CAR-specific mRNA and protein was analyzed by reverse transcriptase polymerase chain reaction and Western blotting, respectively. Adenovirusbased gene delivery was evaluated by infection of cells with adenoviral vector carrying the green fluorescent protein （GFP） gene.RESULTS： All the colon cancer cell lines examined （HT29, LS180, SW480, SW948 and SWlll6） expressed CAR full-length mRNA and an alternatively-spliced variant that lacks the transmembrane coding exon. All cell lines were detected as CAR-positive by Western blot analysis. Further, all cells we examined were efficiently infected with adenoviral vector-GFP.CONCLUSION： The data indicated that the five colon cancer cell lines tested expressed adenovirus primary receptor and could be efficiently infected by adenoviral vectors. Therefore, these cell lines will be useful for adenovirus-based gene transfer and research.展开更多
Menstrual blood stem cells (MensSCs) have enormous potential as a source for cell replacement therapies. Since there is a major concern in utilization of nanofibers in tissue engineering of stem cells, we examined the...Menstrual blood stem cells (MensSCs) have enormous potential as a source for cell replacement therapies. Since there is a major concern in utilization of nanofibers in tissue engineering of stem cells, we examined the potential of MensSCs to differentiate into hepatocytes, using different protocols and compare cells, with two-dimensional (2D) and three-dimensional (3D) culture systems. Cell characterization experiments of MensSCs have demonstrated that they are multipotent stem cells similar to mesenchymal stem cells, which can successfully differentiate into osteogenic and adipogenic lineages. The efficiency of the cells on the scaffold was appraised by scanning electron microscopy (SEM), MTT assay, and hematoxylin and eosin (H&E) staining. Thereafter, the differentiation protocols were developed by hepatocyte growth factor (HGF) and oncostatin M (OSM) with serum-supplemented or serum-free culture media up to 30 days. Immunofluorescence analysis and ELISA assay revealed the expression of albumin (ALB) in differentiated cells. Hepatocyte-like cells expressed liver-specific gene such as albumin(ALB), α-fetoprotein (AFP), tyrosine aminotransferase (TAT) and cytochrome P450 subunit 7a1 (Cyp7a1) at mRNA levels. In conclusion, the evidences presented in this study show that the nanofiber scaffold and MensSCs may provide a source of differentiated cells for treatment of liver diseases.展开更多
Objective:To determine the expression of the synaptonemal complex protein-3 (SYCP3) gene in men with nonobstructive azoospermia. Design:Cross-sectional case study. Setting:Avesina Infertility Clinic,Tehran,Iran. Patie...Objective:To determine the expression of the synaptonemal complex protein-3 (SYCP3) gene in men with nonobstructive azoospermia. Design:Cross-sectional case study. Setting:Avesina Infertility Clinic,Tehran,Iran. Patient(s):One hundred and ten consecutive infertile men presenting nonobstructive azoospermia. Intervention(s):Testicular biopsies for histopathological assessment and analyses of SYCP3 expression level by semiquantitative nested reverse transcription-polymerase chain reaction (RT-PCR). The SYCP3 levels were normalized to expression of the housekeeping phosphoglucomutase 1 gene. Main Outcome Measure(s):Expression of SYCP3 messenger ribonucleic acid (mRNA). Correlation of the histopathological findings with SYCP3 expression levels. Results(s):Testicular SYCP3 mRNA expression was observed in 67/110 (60.9%) patients. The expression level correlated with the degree of spermatogenic failure. Although it was expressed in patients with spermatogenesis and maturation arrest,a lack of expression was seen in all of those men with spermatogonial arrest,Sertoli cell-only syndrome,and testicular atrophy. Conclusion(s):These data indicate that SYCP3 is expressed in human testis and is restricted to germ cells. Our findings,in association with those obtained in experimental animals,shows that lack of SYCP3 expression in human testis may have a negative effect on spermatogenesis and male fertility.展开更多
基金Supported by the Funds from Medical Sciences/University of Tehran and Iranian Ministry of Health and Medical Education
文摘AIM： To investigate the expression of coxsackievirus and adenovirus receptor （CAR） and adenovirus-mediated reporter gene transfer in five human colon cancer cell lines.METHODS： Expression of CAR-specific mRNA and protein was analyzed by reverse transcriptase polymerase chain reaction and Western blotting, respectively. Adenovirusbased gene delivery was evaluated by infection of cells with adenoviral vector carrying the green fluorescent protein （GFP） gene.RESULTS： All the colon cancer cell lines examined （HT29, LS180, SW480, SW948 and SWlll6） expressed CAR full-length mRNA and an alternatively-spliced variant that lacks the transmembrane coding exon. All cell lines were detected as CAR-positive by Western blot analysis. Further, all cells we examined were efficiently infected with adenoviral vector-GFP.CONCLUSION： The data indicated that the five colon cancer cell lines tested expressed adenovirus primary receptor and could be efficiently infected by adenoviral vectors. Therefore, these cell lines will be useful for adenovirus-based gene transfer and research.
文摘Menstrual blood stem cells (MensSCs) have enormous potential as a source for cell replacement therapies. Since there is a major concern in utilization of nanofibers in tissue engineering of stem cells, we examined the potential of MensSCs to differentiate into hepatocytes, using different protocols and compare cells, with two-dimensional (2D) and three-dimensional (3D) culture systems. Cell characterization experiments of MensSCs have demonstrated that they are multipotent stem cells similar to mesenchymal stem cells, which can successfully differentiate into osteogenic and adipogenic lineages. The efficiency of the cells on the scaffold was appraised by scanning electron microscopy (SEM), MTT assay, and hematoxylin and eosin (H&E) staining. Thereafter, the differentiation protocols were developed by hepatocyte growth factor (HGF) and oncostatin M (OSM) with serum-supplemented or serum-free culture media up to 30 days. Immunofluorescence analysis and ELISA assay revealed the expression of albumin (ALB) in differentiated cells. Hepatocyte-like cells expressed liver-specific gene such as albumin(ALB), α-fetoprotein (AFP), tyrosine aminotransferase (TAT) and cytochrome P450 subunit 7a1 (Cyp7a1) at mRNA levels. In conclusion, the evidences presented in this study show that the nanofiber scaffold and MensSCs may provide a source of differentiated cells for treatment of liver diseases.
文摘Objective:To determine the expression of the synaptonemal complex protein-3 (SYCP3) gene in men with nonobstructive azoospermia. Design:Cross-sectional case study. Setting:Avesina Infertility Clinic,Tehran,Iran. Patient(s):One hundred and ten consecutive infertile men presenting nonobstructive azoospermia. Intervention(s):Testicular biopsies for histopathological assessment and analyses of SYCP3 expression level by semiquantitative nested reverse transcription-polymerase chain reaction (RT-PCR). The SYCP3 levels were normalized to expression of the housekeeping phosphoglucomutase 1 gene. Main Outcome Measure(s):Expression of SYCP3 messenger ribonucleic acid (mRNA). Correlation of the histopathological findings with SYCP3 expression levels. Results(s):Testicular SYCP3 mRNA expression was observed in 67/110 (60.9%) patients. The expression level correlated with the degree of spermatogenic failure. Although it was expressed in patients with spermatogenesis and maturation arrest,a lack of expression was seen in all of those men with spermatogonial arrest,Sertoli cell-only syndrome,and testicular atrophy. Conclusion(s):These data indicate that SYCP3 is expressed in human testis and is restricted to germ cells. Our findings,in association with those obtained in experimental animals,shows that lack of SYCP3 expression in human testis may have a negative effect on spermatogenesis and male fertility.
