Objective: Mesenchymal stromal cells (MSCs) are considered as an excellent source in regenerative medicine, but availability and ethical problems limited their routine use. Therefore, another available source with easy procedure and exempt from ethical debate is important. The purpose of this study is to isolate and characterize the MSCs from human placenta. The stromal cells were isolated from Placental Decidua Basalis (PDB-MSC), Umbilical cord Wharton’s Jelly (WJ-MSC) and Amniotic Membrane (AM-MSC).
Methods: Full term human placentas (n=4), from cesarean section delivery were collected. Small fragments from different parts were cultures as explants. The immunophenotyping, mesodermal differentiation, growth kinetics and stemness gene expression was studied.
Results: The cultivated cells from three sources expressed CD44, CD105, and CD90. Gene expression of NANOG and OCT4 confirmed the undifferentiated state. The doubling-times for WJ-MSCs, PLC-MSCs and AM-MSCs, respectively, were 21±8h, 28±9h and 25±9h at passage three and 30±5h, 45±7h and 45±7h at passage tenth. The proliferative potential of WJ-MSCs tended to be higher than the other two sources.
Conclusion: The fetal derives stromal cells; especially the early passages of WJ-MSCs are available supplies for large scale production of MSC for using in clinical studies or research projects.
Objectives: Tacrolimus is widely used as an immunosuppressive drug in liver transplant recipients with a narrow therapeutic range and variable individualized pharmacokinetics. Tacrolimus is a substrate of cytochrome P-450 3A enzyme and the drug transporter, P-glycoprotein. Materials and Methods: We determined the genotypic frequencies of cytochrome P-4503A5 (rs776746), and ABCB1 (rs1045642), single nucleotide polymorphisms in a population of 100 Iranian liver transplant patients, and investigated the influence of the above-mentioned single nucleotide polymorphisms on tacrolimus concentrations. At 7 and 30 days after transplant, tacrolimus dosages (mg/kg/d), trough blood levels (T0), and dose-adjusted concentrations (concentration/dosage ratio) were determined. Polymerase chain reaction, followed by restriction fragment length polymorphism analysis, was used for genotyping cytochrome P-4503A5*3 [6986A>G] as well as ABCB1 [3435C>T]. Results: Ninety-five percent of the population showed a cytochrome P-4503A5*3/*3 genotype. ABCB13435TT genotype was observed in 33 cases (33%); whereas 51 cases (51%) carried 3435CT, and 16 cases (16%) carried 3435CC. With regard to the ABCB1 and cytochrome P-4503A5, they showed no influence on tacrolimus dosing requirements at 1 week or 1 month after transplant. No association of any genetic variant with the acute rejection rate was found. Conclusions: Finally, as the liver donor genotype influences tacrolimus pharmacokinetics with regard to expression of cytochrome P-4503A5, far more than the genotype of the recipient; therefore, it should be considered before recommending any personal immunosuppressive treatment based on pharmacogenetics.
The 14-bp polymorphism in exon 8 of the HLA-G gene is associated with HLA-G mRNA stability and the patterns of alternative isoform splicing and may influence the functionality of the HLA-G molecule. HLA-G expression was related to allograft acceptance and fewer episodes of acute rejection during heart, kidney and liver-kidney transplantation. In order to determine a possible correlation between the 14-bp insertion/deletion polymorphism and kidney allograft outcome in our population, genomic DNA was isolated from 144 patients who had received isolated kidney allografts. The recipients was divided into two groups, grafts presenting features of rejection group and a non-rejection group, and compared them with a control group of 100 healthy subjects. There was no significant difference in allelic frequencies of 14-bp insertion/deletion polymorphism between normal controls and kidney transplant patients. No significant difference was found between the RG and the NRG regarding the 14-bp genotypes and alleles. Therefore, additional studies with more sample size from other populations with analysis of other HLA-G polymorphisms are necessary to define this polymorphism as a valuable clinical marker.
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