ABSTRACT
BACKGROUND: Exosomes have the function of some mesenchymal stem cells. Understanding the substance composition that plays a representative role in mesenchymal stem cell exosomes will provide clues for further exploration of synthetic exosome analogues. OBJECTIVE: To investigate the difference of microRNA expression profiles in exosomes derived from passage 2 and 5 human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS: Exosomes in the supernatant of passage 2 and 5 hUC-MSCs were extracted by ultra-high speed centrifugation. The established library was sequenced by using high-throughput sequencing technology. Then we analyzed the sequence results so as to understand the microRNA expression between different groups, and finally did a cluster analysis. RESULTS AND CONCLUSION: 427 657 kinds of microRNAs were detected in the exosomes from passage 2 hUC-MSCs, accounting for 68.93% of the total microRNAs detected; and 119283 microRNAs were detected in the exosomes from passage 5 hUC-MSCs, accounting for 19.22% of the total microRNAs detected. There were 73 526 microRNAs shared between the exosomes from passage 2 and passage 5 hUC-MSCs, accounting for 11.85% of the total microRNAs detected. Bioinformatics analysis (cluster analysis) results showed that these miRNAs were likely to be involved in 161 biological processes, including cell repair, immune and anti-aging. The microRNAs in exosomes from passage 2 to passage 5 hUC-MSCs were largely different. Partial miRNAs exhibited significantly reduced copy numbers. The top five microRNAs with a higher amount, including has-miR-146a-5p, has-miR-191-5p, has-miR-493-3p, has-miR-423-5p, and has-miR-134-5p, have the potential to be the component of synthetic exosome analogues.
ABSTRACT
BACKGROUND: Studies have shown increasing risks and problems in the serum culture system, such as immune rejection, batch differences and virus risk. In addition, with the discovery and application of exosomes, the serum-free culture system is becoming an increasing concern. OBJECTIVE: To compare the similarities and differences between the serum-free culture system and the traditional serum culture system, which lays the foundation for the clinical transformation of human umbilical cord mesenchymal stem cells (hUCMSCs) and provides experimental data. METHODS: Umbilical cord was collected from term infants of cesarean section under aseptic condition, and hUCMSCs were isolated and cultured by explant tissue technique. hUCMSCs was cultured with 10% fetal bovine serum (FBS) and 15% serum substitutes (AGS) from the original generation. Then an inverted microscope was used to observe cell morphological changes. Flow cytometry was used to detect cell surface markers. Cell counting kit-8 was used to detect cell proliferation. Induced differentiation experiment was used to detect cell differentiation potential. Western Blot was used to detect the protein levels of oct4, nanog and sox2. RESULTS AND CONCLUSION: Under the inverted microscope, hUCMSCs cultured with AGS showed more uniform vortex-like growth, and those cultured with FBS gradually appeared with cell differentiation or aging with the increase of cell generations. hUCMSCs cultured by both methods expressed CD73,CD90 and CD105 but lowly expressed CD34 and CD45, and there was no significant difference between the two culture methods. FBS method was superior to AGS method in proliferation ability. Results from the induced differentiation experiments showed that hUCMSCs cultured by both methods had adipogenic, osteogenic and chondrogenic abilities, and there was no significant difference between the two culture methods. hUCMSC cultured by both methods expressed oct4 and nanog but showed no significant difference in level, while the expression of sox2 was significantly higher in the hUCMSCs cultured by AGS than by FBS (P < 0.05). To conclude, the hUCMSCs cultured with AGS are in accordance with the international standards of mesenchymal stem cells. The AGS method as an alternative to the FBS method can become a preferred method for hUCMSCs culture.
ABSTRACT
BACKGROUND:Studies have shown that the main functions of stem cells include secreting soluble factors,regulating immune balance,suppressing inflammatory response,directional differentiation into specific functional cells involved in tissue repair,promoting revascularization,improving blood circulation and anti-oxidative stress.Therefore,stem cells can be applied for treatment of autoimmune diseases,especially systemic lupus erythematosus.OBJECTIVE:To summarize the immunoregulatory mechanism of mesenchymal stem cells on T and B lymphocytes in systemic lupus erythematosus.METHODS:A computer-based online search of CNKI and PubMed from January 2006 to August 2016 was performed by the first author to search related articles with the keywords of “mesenchymal stem cell,systemic lupus erythematosus,immunoregulation” in Chinese and English,respectively.Articles related to the immunomodulatory effect of mesenchymal stem cells in the treatment of systemic lupus erythematosus were selected,and as for in the same field,literatures published lately in the authoritative journals were preferred.RESULTS AND CONCLUSION:A total of 207 literatures were initially selected,and finally 53 articles were involved in analysis according to the inclusion criteria.Mesenchymal stem cells have the potential of self-renewal and multi-directional differentiation,which are ideal seed cells for immune replacement therapy.Meanwhile,mesenchymal stem cells can treat autoimmune diseases radically because of the ability of immunoregulation.Currently,mesenchymal stem cells have been proved to be a safe and effective therapeutic measure in the treatment of systemic lupus erythematosus,which can avoid drug side effects and have a great clinical prospect.However,there are still numerous problems to be solved in clinic.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the feasibility of HLA haploidentical peripheral blood hematopoietic stem cell transplantation (PBSCT) for patients with β thalassemia major.</p><p><b>METHODS</b>Sixteen patients with β thalassemia major received HLA haploidentical PBSCT from parents. Two conditioning regimens were used. Regimen A was adopted before December 2007, which consisted of fludarabine (total 150 mg/m²), busulfex (total 520 mg/m²), cyclophosphamide (CTX, total 100 mg/kg), antithymocyte globulin (ATG, total 10 mg/kg) and total body irradiation of 3 Gy. Regimen B was adopted after December 2007, which consisted of fludarabine (total 240 mg/m²), busulfex (total 520 mg/m²), CTX (total 100 mg/kg), and ATG (total 10 mg/kg). Combination of cyclosporin (CsA), methotrexate (MTX) and mycophenolate mofetil (MMF) were used for prophylaxis of graft-versus-host disease (GVHD).</p><p><b>RESULTS</b>Of 16 patients, 14 (87.5%) had sustained engraftment. The median days of neutrophil exceeding 0.5 × 10⁹/L and platelet exceeding 20 × 10⁹/L were 13 days (range 10 - 17 days) and 15 days (range 14 - 20 days) after PBSCT, respectively. Complete chimerism was achieved in all the 14 patients at one month after PBSCT. One patient lost his graft with autologous reconstitution 52 days after transplantation. Four patients had grade II-IV acute GVHD and one patient had chronic extensive GVHD. In the 49-month median follow-up duration, 13 of 16 patients were alive in disease-free situation.</p><p><b>CONCLUSION</b>HLA haploidentical PBSCT, which could provide stable and sustained engraftment for thalassemia major patients with no HLA identical donor, is a promising treatment strategy.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Male , HLA Antigens , Genetics , Haploidy , Peripheral Blood Stem Cell Transplantation , Tissue Donors , beta-Thalassemia , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of autologous bone marrow stem cell (BMSC) transplantation via the renal artery on renal function recovery following renal ischemic-reperfusion (I/R) injury in rabbits.</p><p><b>METHODS</b>BMSCs were collected and isolated from rabbits. Twenty-eight rabbits were subjected to renal pedicle clamping for 105 min and randomized subsequently into transplantation group and control group. BMSCs or saline were injected into the kidney via the renal artery, respectively. Before and 1, 3, 5, 7, 14, 21, and 28 days after I/R injury the venous blood was collected to measure the serum levels of SCr and BUN, and the renal tissue was sampled for pathological observation.</p><p><b>RESULTS</b>One and 3 days after I/R injury, serum Cr and BUN levels increased significantly to the highest level in both groups. On the 7th day serum Cr and BUN levels in the transplantation group were lower than those in control group and remained so till the end of the experiment. On the 28th day, the levels of serum Cr (90.1+/-11.1 micromol/L) and BUN (8.0+/-1.5 mmol/L) in the transplantation group were significantly lower than those in the control group (135.6+/-32.5 micromol/L and 10.9+/-2.5 mmol/L, respectively, P<0.05). Pathological observation of the renal tissue revealed renal tubular epithelial cell degeneration, necrosis and abscission.</p><p><b>CONCLUSION</b>BMSC transplantation can accelerate renal function repair after acute tubular necrosis resulting from I/R injury, and decrease serum Cr and BUN levels in early stage following the injury.</p>
Subject(s)
Animals , Female , Male , Rabbits , Acute Kidney Injury , Blood , General Surgery , Blood Urea Nitrogen , Bone Marrow Cells , Cell Biology , Creatine , Blood , Hematopoietic Stem Cell Transplantation , Kidney , Random Allocation , Renal Artery , Reperfusion Injury , Transplantation, AutologousABSTRACT
Objective To investigate whether CD4~+CD25~(high)regulatory T cells(tregs)were increased in the tumor tissue and peripheral blood of prostate cancer patients.Methods The prevalence of CD4~+CD25~(high)regulatory T cells inside the prostate tumor was detected compared with benign tissue from the same prostate.Furthermore,the frequency of CD4~+CD25~(high)regulatory T cells in peripheral blood was detected in prostate cancer patients compared with normal donors.Results Moreover,CD4~+CD25~(high)regulatory T cells from blood and supernatants from cultured prostate tumor tissue samples exhibited immunosuppressive function in vitro.CD4~+CD25~(high)T normal peripheral blood cells were found for the ratio was(0.5?0.1)%.Lower than the proportion of T cells in peripheral blood of patients with CD4~+CD25~(high)was(2.3?0.7)%,P
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of donor-derived NK cells added to pretreatment conditioning regimen on hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.</p><p><b>METHODS</b>Murine model of MHC haplotype-mismatched BMT was established by using BALB/c(H-2d) x C57BL/6(H-2b) (CB6F(1)(H-2d/b)) mouse as recipient, and C57BL/6(H-2b) mouse as donor. Fifty recipient mice were divided into 5 groups. The mice in the first three groups were each infused 1 x 10(6), 5 x 10(5), 2 x 10(5)/mouse donor-derived NK cells, respectively before TBI ((60)Co, 9.0 Gy) and then conditioned with TBI, followed by infusion of C57BL/6(H-2b) mice bone marrow cells four hours later. The mice in the fourth group received TBI only, and in the fifth group, TBI and BMT at the some doses as the first three groups. Hematopoietic reconstitution, survival time, body weight, histopathology of the recipients were followed up.</p><p><b>RESULTS</b>(1) Survival time was (5.15 +/- 0.66) days for the fourth group, and > 30 days for the other 4 groups. (2) Leukocyte and platelet counts at day 10 after BMT were (0.99 +/- 0.22) x 10(9)/L and (61.0 +/- 7.27) x 10(9)/L respectively for the fifth group and (2.01 +/- 0.21) x 10(9)/L, (101.50 +/- 16.34) x 10(9)/L; (1.98 +/- 0.29) x 10(9)/L, (99.50 +/- 16.41) x 10(9)/L and (1.97 +/- 0.21) x 10(9)/L, (98.0 +/- 16.19) x 10(9)/L for the first three groups, respectively. Histopathology displayed no GVHD in all the groups.</p><p><b>CONCLUSION</b>Donor-derived NK cells could promote hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.</p>