ObjectiveTo observe the effect of interleukin (IL) 10 modified endothelial progenitor cells (EPC) in diabetic retinopathy (DR).
MethodsEPC cells were collected and cultivated from the bone marrow of rats and identified by immuno-fluorescence staining. EPC cells were infected with lentivirus (LV) of EPC-LV-IL10-GFP (EPC-LV-IL10-GFP group) or EPC-LV-NC-GFP (GFP group). EPC cells without lentivirus infection was the EPC group. Enzyme-linked immuno sorbent assay (ELISA) was used to measure the concentrations of tumor necrosis factor (TNF)-α, IL10, IL8 and vascular endothelial growth factor (VEGF) in the supernatant of these three groups. 168 male Wistar rats were divided into normal control group (28 rats), diabetes mellitus (DM) group (28 rats), DM-blank control group (56 rats) and DM-intervention group (56 rats). DM was introduced in the latter 3 groups by streptozotocin intravenous injection. Three months later, the rats in the DM-blank control group and DM-intervention group were injected with EPC-LV-NC-GFP or EPC-LV-IL10-GFP by tail vein, respectively. Immunohistochemistry was used to observe the GFP expression in rat retinas. The blood-retinal barrier breakdown was detected by Evans blue (EB) dye. The retinal histopathologic changes were observed by transmission electron microscope. The mRNA level of VEGF, matrix metallproteinases-9 (MMP-9), angiopoietin-1 (Ang-1), inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in retina were measured by reverse transcription-polymerase chain reaction (RT-PCR).
ResultsELISA showed that the levels of TNF-αand IL8 in the supernatant significantly decreased, while the levels of IL10 and VEGF increased (P < 0.05) in EPC-LV-IL10-GFP group. GFP expressed in the retina of blank control group and intervention group, mainly in the ganglion cell layer, inner nuclear layer and outer plexiform layer. The retinal blood vessel pathological change and EB permeability significantly decreased in intervention group compared with DM group (P < 0.05), and blank control group (P < 0.05). RT-PCR revealed that the mRNA level of VEGF, MMP-9 and Ang-1 significantly increased, and eNOS decreased in DM group compared to the normal control group (P < 0.05). The mRNA level of VEGF and iNOS decreased, eNOS increased while Ang-1 and MMP-9 had not changed in DM-blank control group and DM-intervention group compared with DM group (P < 0.05).
ConclusionsIL10 modified EPC can improve the inflammative microenvironment and suppressed the pathogenesis of DR. Furthermore, EPC transplantation can increase the number of EPC and exerted their effect.
Objective To investigate the expression of ATP-binding cassette superfamily G member 2 （ABCG2） in liver cancer cell lines and the relationship between ABCG2 expression and liver cancer drug resistance，and observe the difference of function between the ABCG2 positive cells and negative cells．Methods The expressions of ABCG2 in four liver cancer cell lines （PLC/PRF/5，7402，7701，and 7721） were detected by flow cytometry．IC50 of 5-fluorouracil （5-FU） and adriamycin were calculated．The expression of the ABCG2 in the 7721 cell lines was observed by immunofluorescence staining．The ABCG2 positive and negative cells were selected by the axenic flow sorting method，the difference of function between the ABCG2 positive cells and negative cells was compared．Results The positive expression rate of ABCG2 in the cell line 7721 was highest among four liver cancer cell lines（P＜0.05），and the ABCG2 positive and negative cells had clear bimodal．IC50 of 5-FU and adriamycin to the cell line 7721 were higher than those of the other three cell lines （P＜0.05） except for 5-FU to the cell line 7701．There was no difference in cell proliferation between ABCG2 positive cells and negative cells．Cell cycle analysis showed that the ABCG2 positive cells had more quiescent cells as compared with the negative cells（P＜0.05）．Conclusions ABCG2 expresses in a variety human liver cancer cell lines，ABCG2 positive cells have some stem cell features like drug resistance and more quiescent cells comparing with the negative cells．
Objective To review the application advancements of ATP-binding cassette (ABC) transporter in medical research.Methods Relevant literatures about the applications of ABC families in medical research were reviewed. Results ABC families mainly took roles in transporting substances across cell membrane. Some of them were useful for the prediction of drug resistance and the prognosis of malignant tumors. Others were target s for molecular researches. Their expressions or mutations might be related with the occurrence of diseases. Conclusion ABC families are very important in the diagnosis and therapy for diseases. Thus they are very promising tools for future medical research.
Objective To compare the effectiveness of autologous implantation between bone marrow stem cells and peripheral blood stem cells for treatment of lower limb ischemia. Methods From December 2004 to December 2005, 42 patients with unilateral lower limb ischemia were treated with both autologous bone marrow stem cell implantation(group A, n=21)and autologous peripheral blood stem cell implantation (group B, n=21). Fortytwo patients included 32 males and 10 females. The age ranged from 34 to 80 years, with a mean of 65.6 years. Of the patients, there were 28, 8 and 6 patients suffered from diabetic lower limb ischemia, Burger’s disese and atherosclotic occlusion, respectively. Ischemic history was from 3 months to 5 years, with amean of 2.1 years. A series of subjective indexes (such as improvement of pain, cold sensation and numbness) and objective indexes such as increase of ankle brachial index (ABI), transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing, were used to evaluate the effect. Results After 4 weeks of implantation, the rate of pain relief was 88.2% in group A and 89.5% in group B (Pgt;0.05) ; the rate of cold sensation relief was 94.4% in group A and 94.7% in group B（Pgt;0.05）; improvement of numbness was 69.2% and 66.7% respectively in groups A and B（Pgt;0.05）. Increaseof ABI was 38.1% in group A and 33.3% in group B（Pgt;0.05）; increase of TcPO2 was 85.7% and 90.5% respectively in groups A and B（Pgt;0.05）; angiography was performed in 12 patients (group A) and 9patients (group B), and the new formed collateral vessel rate was 83.3% in group A and 77.8% in group B（Pgt;0.05）; the amputation rate was 9.1% in groups A and B（Pgt;0.05）; the rate of improvement of foot wound healing was 60.0% in group A and 66.7% in group B(P＞0.05). Forty patients were followed up 3-15 months(mean 8 months). The improvement rate of subjective symptoms was 75.0% in group A and 70.0% in group B (Pgt;0.05); increase of ABI was 60.0% in group A and 65.0% in group B; increase of TcPO2 was 80.0% and 75.0% respectively in groups A and B; the new formed collateral vessel rate was 90.0% in group A and 84.6% in group B. All ulcers healed except 1 case in group B. Conclusion Bone marrow stem cell graft and peripheral blood stem cell graft are all effective in treatingower limb ischemia.
Objective To review the value of imaging assessment of stem cell transplantation in treatment for liver cirrhosis.Methods The related literatures in recent years were collected,and the applications of different radiological techniques and strategies of stem cell transplantation in treatment for liver cirrhosis were summarized.Results Stem cell transplantation in treatment for liver cirrhosis was feasible and effective. Radiological assessment could supply the prompt and accurate information for clinic to choose the proper therapeutic method.The curative effect could also be accurately assessed by radiological techniques.Conclusion Radiological examination is important for the assessment of stem cell transplantation in treatment for liver cirrhosis.
Purpose
To investigate the development of embryonic stem cells (ESC)in the subretinal space．
Methods
ESC were cultivated in suspension for 4 days till they developed into cell aggregates,i.e.embryonic body(EB).ESC as well as EB combined with or without RA were respectively transplanted into vitreous cavity and subretina1 space in SD rats,and the subretinal transplanted eyes,transient ischemia-reperfusion injuries were made by ligating the ophthalmic artery for 40 seconds before the transplantation .The experimental eyes were enucleated for histological and immunohistochemical assays after 14～28 d.
Results
The EB was found to develope into photoreceptors induced by RA in the subretinal space under an ischemia-reperfusion condition，and EB transplantation without RA induction induced multiple differentiations in the subretinal space.The single injection of RA without EB induced hyperplasia of the neural retinal cells.ESC transplanted into vitreous cavity rapidly proliferated and developed into atypical hyperplastic mass.
Conclusion
EB derived from ESC can differentiate into photoreceptors induced by RA in the host subretinal space under an ischemia-reperfusion condition.
(Chin J Ocul Fundus Dis，2000，16：213-284)
Objective To investigate the latest development of tissue engineeredregenerative medicine in industrialization, with the intention to direct work in practical area. Methods A complete insight of regenerative medicine in industrialization was obtained through referring to update publications, visiting related websites, as well as learning from practical experience. Results The aerial view of the future of regenerative medicine was got based on knowledge of four different tissue engineering projects. Conclusion All present efforts should be devoted to regenerative medicine area meeting the industrialized trends.
Objective To review the advance in the experimental studies of microRNA(miRNA) and the relationship between miRNA and stem cells. Methods The related literature was reviewed, and the research findings of miRNA and stem cell were summarized. Results miRNA was noncoding small RNA (20-25 nt) involved in posttranscriptional change, that have been shown to regulate gene expressions. Ithas been reported that some kinds of miRNAs were likely important regulators forstem cells maintaining their state of selfrenewal，and play key roles in theirdifferentiation. Conclusion miRNA as regulation of gene expressions, can be served as a new way for stem cells research.
Objective To investigate the experimental condition and mechanism of differentiation of human umbilical cord blood derived mesenchymal stem cells（hUCB-MSC）into neuron-like cells induced by recombined human epidermal growth factor （rhEGF） and taurine in vitro.Methods hUCB-MSC were primary cultured in Dulbeccoprime;s modified Eagle's medium/F12 （DMEM/F-12）which supplemented with 105U/L penicillin G, 100 mg/L streptomycin sulfate, 10% fetal bovine serum (FBS),5% autologous plasma,4 mmol Lglutamine, 30 ng/ml rhEGF.The DMEM/F-12 medium was replaced by taurine medium after 3 passages.The expression of surface antigen CD90,CD29,CD34,CD44 and CD45 were detected by flow cytometry；the expression of neuron specific enolase,rhodopsin and nestin were investigated by immunocytochemistry. The statistical method was chi square test.Results Morphologically similar to bonemarrow MSC,hUCB-MSC became attached cells after the first 5 to 7 days in culture，and reached 80% to 90% confluent after 3 to 4 weeks. Growth accelerated after passage. hUCB-MSC were positive for CD29,CD44 and CD90 but negative for CD34 and CD45. After taurine induction, 2515/3120 cells expressed NSE, 1168/3175 cells expressed rhodopsin and 903/3050 cells expressed nestin while only 234/2965 cells expressed NSE in the control group(P＜0.01).Conclusion rhEGF and taurine can induce hUCB-MSC differentiating into neuronlike or rhodopsin positive cells.
ObjectiveTo study the external biocompatibility bewteen the mouse induced pluripotent stem cells (miPSCs) and poly-3-hydroxybutyrate-co-3-hydroxyhexanoate (PHBHHx).
MethodsAfter we recovered and subcultured miPSCs, we divided them into two groups. There was one group cultured with material of PHBHHx films outside the body. We observed the adhesive pattern of miPSCs on film by fluorescence of 4, 6-diamidino-2-phenylindole (DAPI) staining. The cell vitality was detected by cell counting kit-8 (CCK-8). The morphology of miPSCs attached on the film was visualized under scanning electron microscope (SEM). We used the traditional petri dish to culture miPSCs and detect the cell activity by CCK-8.
ResultsMiPSCs can adhere and proliferate on PHBHHx films. The result of cell vitality which detected by CCK-8 showed that there was a statistical difference in OD value between culturing on PHBHHx films and traditional cultivation (0.617±0.019 vs. 0.312±0.004, P < 0.05).
ConclusionThere are adhesion and proliferation on the surface of cells patch made by miPSCs co-culturing with PHBHHx film. Compared with traditional culturing in the cell culture dish, culturing in PHBHHx films have great advantages in the process of adhesion and proliferation. PHBHHx can be used as one of the scaffold for stem cells treating various disease.
