Immunoregulatory Effect of Adipose Mesenchymal Stem Cells on Peripheral Blood Lymphocytes in Psoriasis Vulgaris Patients / 中国医学科学院学报

Objective To investigate the effect of adipose mesenchymal stem cells(AMSCs) on the peripheral blood lymphocyte(PBL) in psoriasis vulgaris(PV) patients and the expression and secretion profiles of related inflammatory cytokines in the PBL.Methods AMSCs from three PV patients were co-cultured with PBL. Peripheral blood regulatory cells(Treg) and T helper cell 17(Th17)ratio was measured by flow cytometry. The anti- and pro-inflammatory cytokines expressed and secreted by PBL were detected by quantitative real-time polymerase chain reaction(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA).Results The Treg/total lymphocyte ratio was significantly higher in the healthy people AMSCs+PBL co-culture group[(3.2±0.5)%;P=0.001],but AMSCs in patients had a tendency to promote the proliferation of Treg cells [(1.3±0.2)%],with no significant difference(P=0.485) when compared with the PBL culture alone group[(1.0±0.1)%]. qRT-PCR showed that the ability of PBL in expressing Treg transcription factor forkhead box p3 and transforming growth factor(TGF)-Β mRNA was significantly lower in psoriasis AMSCs+PBL co-culture group than in the healthy people AMSCs+PBL co-culture group(P=0.00,P=0.03),AMSCs had a tendency to promote the expression of interlukin(IL)-10 in peripheral blood lymphocytes,but there was no significant difference(P=0.09).ELISA showed the PBL in healthy people AMSCs+PBL co-culture group secreted the anti-inflammatory cytokine IL-10[(156.9±41.8) ng/Μl] and TGF-Β[(2774.1 ± 526.4) ng/Μl];in contrast,the abilities of PBL in PV patient AMSCs+PBL co-culture group in secreting the anti-inflammatory cytokines has a downward trend:IL-10[(90.4±28.8) ng/Μl] and TGF-Β[(1597.9±55.7) ng/Μl],although the differences were not statistically significant. After the co-culture,the proportion of Th17 cells in the psoriasis AMSCs+PBL co-culture group[(0.8±0.3)%] showed a decreasing trend when compared with the PBL culture alone group[(1.1±0.1)%],although the results were not statistically significant. Also,the proportion of Th17 cells showed no significant difference between PV patient AMSCs+PBL co-culture group and healthy people AMSCs+PBL co-culture group. Finally,both the psoriasis AMSCs+PBL co-culture group and the healthy people AMSCs+PBL co-culture group showed no obvious inhibitory effect on the expression and secretion of Th17 transcription factor retinoid-related orphan nuclear receptor Γt and pro-inflammatory cytokines IL-17 and IL-23 in PBL,and there was no significant difference between these two groups.Conclusions AMSCs in PV patients have decreased ability in regulating the anti-inflammatory function of peripheral blood Treg lymphocytes. However,they have no effect on the proinflammatory effect of peripheral blood Th17 lymphocytes.

miR-210 plays a role in the transformation from mesenchymal stem cells (MSCs) into cancer-associated fibroblasts (CAFs) / 基础医学与临床

Objective To investigate whether miR-210 plays a crucial role in the process of inducing the transfor-mation from co-cultured adipose derived(AD)-MSCs into CAFs. Methods Co-cultured adipose derived (AD)-MSCs with breast cancer cells (BCCs) lines in vitro and detected the expression of miR-210 using RT-qPCR. The expression of CAFs characteristic markers including α-SMA and tenascin-c were measured by RT-qPCR and the ex-pression of α-SMA and FAPA were assessed by Western blot.Co-injected MCF-7 cells transfected with miR-210 in-hibitor or miR-NC with MSC at 1 : 1 ratio into the immunodeficient nude mice subcutaneously and observed tumor growth in vivo constantly. Results miR-210 was all up-regulated when a variety of breast cancer cells were co-cul-tured with MSCs (P<0.05). Inhibition of miR-210 in tumor cell could inhibit the transformation from MSCs into CAFs(P<0.05). Animal experiments further confirmed that inhibition of miR-210 in tumor cell could reduce tumor growth (P<0.05). Conclusions As an important information molecule, miR-210 mediates the transforma-tion from MSCs to CAFs in the tumor microenvironment.

Effects of Wnt3a and Wnt5a signaling pathway switching on hematopoietic stem cells senescence / 基础医学与临床

Hematopoietic stem cells(HSCs)senescence is an important reason of hematopoietical and immunologi-cal function senescence.It also is play a key role during aging-related diseases development.Under certain condi-tions,the activation of classical Wnt 3a/β-catenin is in favour of maintains polarity and young states of HSCs,self-renewing,proliferation and differentiation potency.Switching to the non-classical Wnt5a pathway,further activation of Cdc42 protein and others can promote HSCs ageing,and indirectly inhibits Wnt3a/beta-catenin pathway.The in-tervention of two Wnt signaling pathways switching and mechanism,not only can illustrate the mechanism of HSCs aging,but also clear how to slow down ageing.This could provide a new strategy on the solution of age-related dis-eases and keeping a young state.

Establishment of stem cell curriculum and thinking of teaching practice in graduate education / 医学研究生学报

In order to provide reference for development of the new curriculum for graduate students "The principle,technical and clinical practice of stem cell" and improve the quality of teaching and learning,we summarized some experiences and lessons for this course.This paper introduced some measures and achievements on the teaching reform from several aspects including curriculum design,selection of teaching content,teaching idea and mode,training the teaching groups and so on.Moreover,it especially emphasized that teaching contents should not only pay attention to the basic and clinical application,but also combine the present laws and keep abreast of scientific and technological developments.In the process of teaching,we need to value the quality of teachers and improve teaching standards,meanwhile we should pay attention to the culture of thought and the capacity of innovation,stimulating students' interests in studies,improving autonomous study and practices ability through multiple means and channels.This course teaching got satisfactory results and the acquired practice experiences were worth of being popularized in the further graduate students teaching.

Protection of Nerve Injury with Exosome Extracted from Mesenchymal Stem Cell / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the protective effect of Exosomes from human adipose-derived mesenchymal stem cells (hAMSCs) in neural injury induced by glutamate and its possible mechanism.</p><p><b>METHODS</b>Characteristics of Exosomes from hAMSCs were identified by electron microscopy and Western blot analysis. Cytokines that might play a major role in the protective effect were tested by enzyme-linked immunosorbent assay (ELISA). The protective action of Exosome and its possible signaling pathway were researched by the in vitro neural injury induced by glutamate, including control group (without Glu), Glu group (dealing with Glu), Glu+Exo group (dealing with Glu +100 ng/ml Exo), Glu+Exo+Akt group (dealing with Glu+100 ng/ml Exo+10 μmol/L Akt), Glu+Exo+Erk group (dealing with 100 ng/ml Glu+100 ng/ml Exo+10 μmol/L Erk), and Glu+Exo+TrkB group (dealing with Glu+100 ng/ml Exo +10 μmol/L TrkB).</p><p><b>RESULTS</b>Exosomes from hAMSCs had similar sizes to those isolated from other kinds of cells, and expressed the characteristic proteins such as CD63, CD81, HSP70, and HSP90. Cytokines that had neurotrophic effects on Exosomes were mainly insulin-like growth factor and hepatocyte growth factor, with the concentration being 9336.49±258.63 and 58,645.50±16,014.62, respectively; brain derived neurotrophic factor, nerve growth factor,and vascular endothelial growth factor had lower levels, with the concentration being 1928.25±385.47, 1136.94±5.99, and 33.34±9.43, respectively. MTS assay showed that the PC12 cell survival rates were 0.842±0.047, 0.306±0.024, 0.566±0.026, 0.461±0.016, 0.497±0.003, and 0.515±0.034 in the control group, Glu group, Glu+Exo group, Glu+Exo+Akt group, Glu+Exo+Erk group, and Glu+Exo+TrkB group; obviously, it was significantly lower in Glu group than in control group (P=0.02), significantly higher in Glu+Exo group than in Glu group (P=0.01), and significantly lower in Glu+Exo+Akt group than in Glu+Exo group (P=0.01).</p><p><b>CONCLUSION</b>Exosomes secreted from hAMSCs have protective effect against neuron damage induced by glutamate, which may be mediated through activating the PI3/K-Akt signalling pathway.</p>

Concept of mesenchymal stem cells: bring more insights into functional research of MSC / 中国实验血液学杂志

Mesenchymal stem cells have generated great interest among researchers and physicians due to their unique biological characteristics and potential clinical applications. Here, I propose for the first time the concept of a hierarchical system which is composed of all mesenchymal stem cells from post-embryonic subtotipotent stem cells to MSC progenitors. Post-embryonic subtotipotent stem cells are left-over cells during embryonic development and are on the top of the hierarchy. MSC system is a combination of cells that are derived from different stages of embryonic development, possess different differentiation potential and ultimately give rise to cells that share a similar set of phenotypic markers. The concept of MSC system has important implications: (1) it entirely explains the three important biological characteristics of MSC: stem cell properties of MSC, MSC as components of tissue microenvironment and immunomodulatory functions of MSC. (2) It balances immune responses and tissue metabolism. (3) It could provide tissue-specific stem cells for clinical application with high efficiency and safety. In a word, this concept constitutes an important part of the biological properties of MSC and will help researchers gain better insight into MSC.

Study on inducing differentiation of human amniotic epithelial cells into insulin secreting cells in vitro / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the differentiation of human amniotic epithelial cells (hAECs) into insulin secreting cells (ISCs) in vitro.</p><p><b>METHODS</b>The hAECs were isolated from human amnion by trypsin digestion, and the phenotype of the isolated cells were identified by flow cytometry and immunocytochemical staining. The hAECs at passage 3 were treated with nicotinamide and N2 supplement to investigate their differentiation into ISCs. At different times after differentiation, the expression of insulin and beta2 microglobulin (beta2-MG) was determined by immunocytochemical staining, while the content of insulin in supernatant from cultured hAECs was detected by radioimmunoassay and the expressions of insulin, pancreatic and duodenal homeobox factor-1 (PDX-1) mRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) hAECs expressed high percent of CD29, CD73, CD166 and CK19. (2) At 7, 14 and 21 days, the percentages of insulin-positive cells in induced groups were 74.00% +/- 1.73%, 75.33% +/- 1.15% (see symbol) 75.67% +/- 0.58% respectively, which were negative in control groups. (3) At 7, 14 and 21 days, contents of insulin in supernatant from induced groups were (328.47 +/- 3.22) microIU/ml, (332.26 +/- 1.22) microIU/ml and (329.68 +/- 2.57) microIU/ml respectively, they were significantly higher than those in control groups (All P < 0.01). (4) PDX-1 mRNA and beta2-MG were expressed before and after the induction of hAECs, but insulin mRNA was expressed only in the induced groups.</p><p><b>CONCLUSION</b>hAECs can differentiate into ISCs, having the potential application for therapy of type I diabetes.</p>

Virus free induction of umbilical cord derived mesenchymal stem cells into islet-like cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the feasibility of using a virus-free system in the induction of umbilical cord derived mesenchymal stem cells (UC-MSCs) into insulin-secreting cells.</p><p><b>METHODS</b>MSCs were isolated from human umbilical cord and induced into insulin-secreting cells with a three-stage method. The mRNA expression levels of foxa2, sox17, pdx1, ngn3, pax4, insulin, and glut-2 were compared between induced and non-induced groups by RT-PCR in each stage. The distribution pattern of insulin and c-peptide were detected by immunofluorescence staining and observed by fluorescence microscopy. Insulin and c-peptide secretion and glucose responsiveness were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Transcription factors foxa2, sox17, pdx1, ngn3, pax4, insulin, and glut-2 were expressed in the induced cells. The mRNA expression levels of foxa2 and sox17 were significantly higher in the induced group than those in non-induced group in the first stage (all P < 0.05), pdx1, ngn3, and pax4 were significantly higher in the induced cells than those in non-induced cells in the second stage (all P < 0.05), and insulin and glut-2 expressions were significantly up-regulated in the induced group at last stage (all P < 0.05). Immunofluorescence staining showed that insulin and c-peptide were located in the cytoplasm of more than 90% of induced cells. ELISA showed that total intracellular insulin content of the induced cells contained up to (346.3 739 +/- 32.5 149) microU/ml, which was significantly higher than insulin in non-induced cells (17.69 +/- 1.46) microU/ml (P < 0.01). C-peptide content of the induced cells measured up to (195.10 +/- 8.88) pmol/L/h (P < 0.01), when exposed to 5.5 mmol/L glucose (P < 0.01). When stimulated with 22 mmol/L glucose, the c-peptide content of the induced cells increased to (340.99 +/- 7.91) pmol/L/h (P < 0.01 ).</p><p><b>CONCLUSION</b>The umbilical cord derived MSCs can be efficiently induced into insulin-secreting cells via a virus-free system.</p>

Methylation of Id4 gene and inhibitive effect of arsenic trioxide on it in Raji cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study methylation of Id4 gene and demethylation effect of arsenic trioxide (ATO) in Raji cells.</p><p><b>METHODS</b>Human Burkitt's Raji lymphoma cells were cultared and treated with ATO at different concentrations and different time points. Methylated degree of Id4 gene was detected by methylation specificity polymerase chain reaction (MS-PCR), Id4 mRNA expression in Raji cell by reverse transcription polymerase chain reaction (RT-PCR), the growth of cell by MTT assay, and cell apoptosis and cycle distribution by Flow Cytometry (FCM).</p><p><b>RESULTS</b>(1) The Id4 gene exhaustive methylation in control group, and hypermethylation in experimental group were reversed by ATO in a dose-dependent manner. (2) Id4 mRNA expression in Raji cells treated with ATO for 48 h increased gradually with ATO concentration increasing in experimental group. (3) Raji cell growth inhibited rates after different concentrations of ATO treatment for 24, 48, 72 h were 12.15% ∼ 92.17% in the experimental group (P < 0.05). (4) Apoptosis peak emerged after ATO treatment for 48 hours in experimental group, while a much lower apoptosis in control group. (5) After ATO treatment for 48 h in experiment group, the cells were arrested at G(0)/G(1) phase in a dose-dependent manner.</p><p><b>CONCLUSION</b>Id4 gene presents exhaustive methylation in Raji cells. ATO can reverse the hypermethylation of Id4 gene and recover the expression of Id4 mRNA. Hypermethylation of Id4 gene is one of the reasons of Raji cells malignant proliferations.</p>

Transfusion of mesenchymal stem cells combined with haploidentical HSCT improves hematopoietic microenvironment / 中国实验血液学杂志

The aim of this study was to investigate the role of mesenchymal stem cells in the hematopoietic reconstitution of patients who had received haploidentical allogeneic hematopoietic stem cell transplantation (hi-allo-HSCT). 15 patients who underwent treatment with both MSCs and HSCs, were selected as study group, while 20 patients receiving only HSCT were taken as control. Bone marrow samples were obtained from iliac crest aspirates at several times after HSCT for the isolation, purification and expansion of MSCs. The confluent ratio and time were measured and compared with those of the control. The peripheral blood samples were obtained from patients, then absolute neutrophil and platelet counts were assayed. From day 4 before transplantation to day 28 after transplantation, serum was obtained every four days from patients of the two groups, and then 3 cytokines as SDF-1alpha, TPO and IL-11 were detected by ELISA. The results indicated that as compared with the control group, the ratio of primary confluent layer formation of MSCs in study group was obviously higher (27.3%) (p < 0.01), and the confluence time in culture was significantly less (p < 0.05). In the study group, the concentration of SDF-1alpha amounted to peak value (2975.19 +/- 681.56 pg/ml) on the 8th day after HSCT, which was obviously higher than that before HSCT (2403.70 +/- 522.39 pg/ml, p < 0.05), whereas in the control, the concentration of highest point of SDF-1alpha reached to peak valve (2280.60 +/- 701.25 pg/ml) on the 16th day after HSCT, which was less than that before HSCT (2701.46 +/- 483.21 pg/ml, p < 0.05). The concentration of TPO and IL-11 was higher in study group compared with the control from day 16 to 28 after HSCT (p < 0.05). It is concluded that the transfusion of MSCs combined with hi-all-HSCT may improve the injured state of the hematopoietic microenvironment in bone marrow of patients during allo-HSCT.

Therapeutic efficacy of bone marrow-derived mesenchymal stem cells infused into mice with liver injury induced by concanavalin A / 中国实验血液学杂志

The aim of this study was to evaluate the therapeutic potential of bone marrow mesenchymal stem cells (MSC) on acute liver injury induced by concanavalin A (ConA). MSCs were isolated from male C57BL/6 mice and cultured, and a ConA-induced acute liver injury model was used. MSCs were systemically infused immediately after mice were challenged with ConA, control mice received only saline infusion. 24 hours after MSC transplantation, the level of serum aminotransferases, histologic change and in situ apoptosis of cells were detected, the expression of inflammatory mediators were examined by real-time RT-PCR. The results indicated that MSC transplantation significantly reduced ConA-induced acute liver injury, including the decrease of the level of serum alanine aminotransferase (ALT), serum aspartate aminotransferase (AST) and the extenuation of liver necrosis and in situ apoptosis. Furthermore, after MSC infusion the expression of TNF-alpha, IFN-gamma in liver decreased greatly (p<0.05) with no statistical difference in the expression of iNOS, IL-2 and IL-10 (p>0.05). It is concluded that the systemic infusion of MSCs can alleviate ConA induced acute liver injury in mice.

In vitro study of directional inducible differentiation of inner ear hair cells from human adipose-derived mesenchymal stem cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the feasibility of directionally inducing human adipose-derived mesenchymal stem cells (hAD-MSCs) towards inner ear hair cells.</p><p><b>METHODS</b>Mesenchymal stem cells from human adipose tissue were isolated, purified and cultured in vitro. hAD-MSCs were induced to neural stem/progenitor-like cell, and then co-cultured with embryonic chick otic vesicle cells. Processed hAD-MSCs were tested by immunostaining to ascertain whether they expressed characteristic hair cell markers.</p><p><b>RESULTS</b>Morphologically, hAD-MSCs were induced to differentiate into neural stem/progenitor cells and expressed specific neural markers. After being co-cultured with embryonic chick otic vesicle cells, hAD-MSCs expressed specific surface markers of inner ear hair cells.</p><p><b>CONCLUSIONS</b>hAD-MSCs can be directionally induced to differentiate towards hair cell-like cells in vitro.</p>

Progress of study on the roles of matrix metalloproteinases in the pathogenesis of hematologic malignancies -- review / 中国实验血液学杂志

Matrix metalloproteinases (MMPs) have the ability to degrade extracellular matrix components. They abnormally express in a variety of solid tumors and hematologic malignancies, and play an important role in tumor invasion and metastasis through extracellular matrix degradation, which closely relates with the progression and prognosis of the malignant disease. This article reviews progress of study on the mechanism of MMP underlying the pathogenesis of hematologic malignancies, including structure of MMP, regulation mechanism of MMP and its relation with proliferation and differentiation of hematopoietic stem cells (HSC), angiogenesis and tumor immunology and so on.

The effect of TNF-alpha on TAZ expression and osteogenic potential of mesenchymal stem cells from myeloma patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the relationship between TNF-alpha, transcriptional co-activator with PDZ-binding motif (TAZ) and bone disease of multiple myeloma.</p><p><b>METHODS</b>The biological characteristics, especially the osteogenic potential of marrow MSCs from myeloma patients and normal subjects were studied. Real-time RT-PCR and Western-blot were employed to detect mRNA and protein expression of TAZ in MSCs. The concentration of TNF-alpha in the marrow plasma was detected using ELISA method. CD138(+) myeloma cells were cocultured with normal MSCs with or without anti-human TNF-alpha monoclonal antibody in the Transwell system. Real-time RT-PCR was employed to detect the mRNA expressions of ALP, Cbfa1 and TAZ in MSCs two weeks later. von Kossa staining was used to detect the mineral deposition. TNF-alpha was added into the culture media of normal marrow MSCs and real-time RT-PCR and Western-blot were employed to detect mRNA and protein expression of TAZ in MSCs one week later.</p><p><b>RESULTS</b>Real-time RT-PCR revealed that the mRNA of osteogenic markers was decreased in comparison with that of normal controls after cultured in the osteogenic medium. von Kossa staining showed weakened mineral deposition in MSCs from multiple myeloma patients compared with that in normal subjects after osteogenic differentiation for two weeks. The mRNA and protein levels of TAZ in the MSCs from myeloma patients were decreased. TNF-alpha concentration in the marrow plasma of myeloma patients was higher than that in the normal controls [(355.4 +/- 49.1) vs. (92.3 +/- 17.2) pg/ml]. CD138(+) myeloma cells inhibited mRNA expressions of ALP, Cbfal1 and TAZ in MSCs, which could be partially reversed by anti-human TNF-alpha monoclonal antibody.</p><p><b>CONCLUSION</b>The osteogenic potential of MSCs from myeloma patients is significantly decreased in comparison with that in normal subjects, which may play an important role in the pathology of myeloma bone disease. TAZ expression inhibited by TNF-alpha may play an important role in this inhibition effect.</p>

Effect of total body irradiation on cellular senescence related indexes of bone marrow mesenchymal stem cells / 中国实验血液学杂志

The aim of this study was to explore the changes in cellular senescence related indexes of bone marrow mesenchymal stem cells (BMMSCs) after total body irradiation (TBI). At different time points after 4 Gy irradiation, BMMSCs were isolated from male C57BL/6 mice and cultured. Morphology, senescence-associated beta-galactosidase (SA-beta-gal) staining and cell cycle analysis were used to evaluate the changes in BMMSCs at cellular level while real-time RT-PCR was used to detect the alterations in senescence related gene expression including p16INK4a, p21Cip1/Waf1, p53 and TGF-beta1. The results showed that within 4 weeks after exposure to 4 Gy TBI, the morphology of BMMSCs and the expression level of SA-beta-gal were not significantly changed, the cellular senescence-related cell cycle arrest was not occurred and the senescence related gene expression level was not increased. It is concluded that at the early stage after 4 Gy TBI, the related molecular level of cellular senescence in BMMSCs is not changed.

Immuno-suppressive effects on T cells mediated by mesenchymal stem cells from patients with myelodysplastic syndrome / 中国实验血液学杂志

The study was aimed to compare the effects of T-cell suppression mediated by mesenchymal stem cells (MSC) from normal individuals and myelodysplastic syndromes (MDS) patients. MSC were cultured from the bone marrow of 12 healthy volunteers and 12 MDS patients, the morphology, surface markers and expression of several cytokines of MSC from normal individuals and MDS patients were compared, and the effects of T-cell suppression were tested in the following assays: phytohemaglutinin (PHA)-primed cultures, mixed lymphocyte reaction (MLR), cell cycle of T-cell after PHA-primed cultures and apoptosis of T-cell as well. The results showed that the MSC from normal individuals and MDS patients were similar in morphology, proliferation and surface markers. The suppressions of T-cell proliferation induced by PHA and alloantigens mediated by MDS-MSC were significantly lower than that of normal MSC. More T-cells were arrested in G0/G1 phase by normal MSC, while the effects were deficient by MDS-MSC. The suppression of T-cell activation mediated by MDS-MSC was also lower than that of normal MSC, but suppression effect on T-cell apoptosis increased. The cytokines TGF-beta1, 3, FasL expressed by MDS-MSC were reduced as compared with normal MSC, but TGF-beta2 expression increased in MDS-MSC. It is concluded that although the morphology, proliferation and cell surface markers of MDS-MSC are normal, the T-cell suppression mediated by MDS-MSC is deficient as compared with normal controls. Whether these abnormalities are relevant to the pathogenesis of aplastic anemia remains to be determined.

Ferumoxide labeled Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells and its in vivo tracing in the brains of Macaca Fascicularis / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the method for labeling Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells (hBMSCs) with ferumoxide-PLL and evaluate the feasibility of its tracing after transplantation into the brains of Macaca Fascicularis.</p><p><b>METHODS</b>The hBMSCs were incubated with ferumoxide-PLL. Trypan blue staining, Prussian blue staining, and transmission electron microscope were performed to show intracellular iron, marking efficiency, and the vigor of the labeled cells. After the hBMSCs were transplanted into the brains of cynomolgus monkeys by stereotaxis, magnetic resonance imaging (MRI) was performed to trace the cells in vivo. Cell survival and differentiation were studied with immunohistochemistry, Prussian blue staining, and HE staining.</p><p><b>RESULTS</b>The marking efficiency of the ferumoxide-PLL was 96%. Iron particles were found intracytoplasmic of the hBMSCs by Prussian blue staining and transmission electron microscopy. The relaxation rates of labeled cells in MRI were 4.4 and 4.2 times higher than those of the unlabeled cells. Hypointensity area was found by MRI three weeks after transplantation. Many hBMSCs and new vessels were found in the transplantation zone by pathological and immunofluorescence methods.</p><p><b>CONCLUSIONS</b>Ferumoxide-PLL can effectively label hBMSCs and thus increase its contrast in MRI results. The cells can survive in the brains of cynomolgus monkeys. The labeled hBMSCs can be traced in vivo by MRI.</p>

Mice adipose derived Flk-1+ mesenchymal stem cells can ameliorate Duchenne's muscular dystrophy in Mdx mice for their multilineage potential / 中国实验血液学杂志

Duchenne muscular dystrophy (DMD) is a common X-linked disease characterized by widespread muscle damage that invariably leads to paralysis and death. There is currently no therapy for this disease. This study was purposed to investigate the feasibility to use adult adipose-derived mesenchymal stem cells (AD-MSCs) in the therapy of DMD. The Flk-1(+) MSCs were isolated from adipose tissue of adult GFP mice; the phenotype and cell cycle of MSCs were analyzed by flow cytometry; the AD-MSCs were directionally differentiated by myoblast and endotheliablast induction system in vitro and were identified by immumofluorecence staining and RT-PCR; the AD-MSCs were transplanted into CTX-injured mice model or mdx mice (DMD animal model) through tail vein; the distribution and differentiation of AD-MSCs were detected by immunofluorescence staining and RT-PCR respectively, and statistic analysis was performed. The results showed that the Flk-1(+) AD-MSCs could be induced to differentiate into myoblasts and endothelial cells in vitro. After transplanted into CTX-injured mice model or mdx mice, GFP-positive cells could be detected in damaged muscle, and these donor-derived cells were also positive for MHC, vWF, or Pax7. Flk-1(+) AD-MSC transplantation also partly reconstituted the expression of dystrophin, and reduced the percentage of centronucleated myofibers in mdx mice. It is concluded that Flk-1(+) AD-MSCs represent a possible tool for future cell therapy applications in DMD disease, as they can be delivered through the circulation for their potential of muscle homing. And Flk-1(+) AD-MSCs also show the ability to contribute to muscle repair, improvement of blood supply and long term reconstitution of dystrophy muscle.

Reduction of bone marrow mesenchymal stem/progenitor cells pool and alteration of their osteogenesis potential caused by total body irradiation / 中国实验血液学杂志

To investigate the effect of irradiation on the quantity and osteogenesis potential of BMMSCs and to explore the response of them in the irradiation stress and its contribution to long-term effects of radiation-induced bone and hematologic injury, a total body irradiation (TBI) murine model was adopted. The number of CFU-F and cell cycle profile of BMMSCs were analyzed at different time points before and after TBI. Osteogenic differentiation was evaluated by Von Kossa staining, expressions of osteogenesis-related genes and transcriptional coactivator with PDZ-binding motif (TAZ) were detected by real-time RT-PCR. The results showed that the number of CFU-F decreased greatly at day 28 after TBI. At day 3 after TBI, more cells entered cell cycle and the osteogenesis potential was greatly enhanced followed by recovery of cell cycle distribution and significant defect in osteoblast differentiation respectively, meanwhile the expression of TAZ was changed. It is concluded that TBI results in the reduction of bone marrow mesenchymal stem/progenitor cell pool and alters the osteogenesis potential of BMMSCs, which is related to the change of TAZ expression.

Effect of hypoxia on mesenchymal stem cells - review / 中国实验血液学杂志

Oxygen is essential for life, but cultivation of cells is usually performed under 20% O(2), that do not replicate normal physiological hypoxia or pathological hypoxia conditions in the body. Recently, the effect of hypoxia on mesenchymal stem cells (MSCs) has been studied, under physiological hypoxia, MSCs thrive well, and the ability differentiating to osteoblast, chondrocyte and adipocyte as well as the ability of migration are changed. Hypoxia changes the physiological characteristics of embryonic stem cell, hematopoietic stem cell and neuron stem cell as well. The mechanism of these responses might be primarily involved in the hypoxic inducible factor-1 (HIF-1) signal pathway. This review emphasizes that hypoxia is an important factor on all major aspects of stem cell biology including survival, proliferation, differentiation, and migration, and the mechanism involved in HIF-1 signaling pathway behind these responses was also discussed.