Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 46
Filter
1.
Article in Chinese | WPRIM | ID: wpr-1020918

ABSTRACT

Objective To investigate the effects of peroxisome proliferator activated receptor-gamma(PPARγ)gene silen-cing in human bone marrow stromal cells(HS-5)on hematopoietic function in bone marrow-suppressed mice,and to explore the potential mechanisms involved.Methods A bone marrow-suppressed mouse model was established by whole-body X-ray irradi-ation.Two hours after modeling,the mice were randomly divided into three groups:experimental group(intravenous injection of PPARγ RNAi-interfered HS-5 cells through the tail vein),control group(intravenous injection of PPARγ RNAi-uninterfered HS-5 cells through the tail vein),and blank group(intravenous injection of an equal amount of saline through the tail vein),with 5 mice in each group.Peripheral blood routine tests were performed before,24 hours after,1 week after,and 2 weeks after radio-therapy.In vitro osteogenic and adipogenic induction was performed in cells,and the cells were divided into experimental group(PPARγ RNAi-interfered HS-5 cells),control group(PPARγ-uninterfered HS-5 cells),and blank group(HS-5 cells without os-teogenic/adipogenic induction).Osteogenic/adipogenic staining was observed.The effects of PPARγ gene-silenced HS-5 cells on mouse bone marrow hematopoietic stem cells(HSCs)were detected by CCK-8 proliferation assay.The groups included experi-mental group(PPARγ RNAi-interfered HS-5 cells were co-cultured with mouse HSCs after 3 days of osteogenic induction dif-ferentiation),positive control group(HS-5 cells treated with 50 μmol/L PPARγ inhibitor were co-cultured with mouse HSCs af-ter 3 days of osteogenic induction differentiation),negative control group(PPARγ RNAi-uninterfered HS-5 cells were co-cul-tured with mouse HSCs after 3 days of osteogenic induction differentiation),and blank group(Mouse HSCs were cultured alone without co-culturing with HS-5 cells).Results After radiotherapy,the hematological parameters of mice in each group showed a decreasing trend initially,and then increased.One week after radiotherapy,there were significant differences in platelet and white blood cell levels among the three groups(experimental group>control group>blank group,all P<0.05).Two weeks after radiotherapy,there were significant differences in the percentage of adipocyte vacuole area among the three groups(experi-mental group<control group<blank group,all P<0.05).Pearson correlation analysis showed a negative correlation between hematological parameters and PPARγ expression levels(all P<0.05),as well as a negative correlation between hematological parameters and the percentage of adipocyte vacuole area(all P<0.05).After in vitro osteogenic/adipogenic induction differenti-ation,compared to the control group,the experimental group showed a significantly lower proportion of orange-red cells and a significantly higher proportion of red calcium nodules.After 3 days of osteogenic induction differentiation,the experimental group,positive control group,and negative control group of human bone marrow stromal cells were co-cultured with mouse HSCs,while HSCs were solely cultured in the blank group.The results showed that after 24 h,48 h and 72 h of co-culture,the A values of mouse HSC cells in the experimental group and positive control group were higher than those in the negative control group and blank group(all P<0.05).Conclusion Silencing of the PPARγ gene in HS-5 cells implanted into bone marrow-sup-pressed mice contributes to enhanced hematopoietic function in mice.After interference and silencing of the PPARγ gene,the os-teogenic differentiation ability of HS-5 cells is enhanced,while the adipogenic differentiation ability is weakened.Furthermore,osteogenic-induced HS-5 cells can further enhance the proliferation capacity of mouse HSCs.

2.
Article in Chinese | WPRIM | ID: wpr-928713

ABSTRACT

Acute lymphoblastic leukemia (ALL) is a kind of the most common hematopoietic malignancy, its recurrence and drug resistance are closely related to the bone marrow microenvironment. Bone marrow stromal cell (BMSC) is an important part of the bone marrow microenvironment and their interaction with leukemia cells cannot be ignored. BMSC participates in and regulate signaling pathways related to proliferation or apoptosis of ALL cells by secretes cytokines or extracellular matrix proteins, thus affecting the survival of ALL cells. In this review, the research advance of several signaling pathways of the interaction between BMSC and ALL cells was summarized briefly.


Subject(s)
Humans , Apoptosis , Bone Marrow , Bone Marrow Cells , Mesenchymal Stem Cells , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Stromal Cells , Tumor Microenvironment
3.
Article in Chinese | WPRIM | ID: wpr-663715

ABSTRACT

Bone marrow stromal cells (BMSCs) are a kind of stem cells with multiple differentiation potential in bone marrow.BMSCs have been widely used in tissue engineering,cell transplantation,gene therapy and organ transplantation,due to their characteristics of wide range of sources,weak immunogenicity weak,easily transfected by exogenous gene,long survival time in the host,multi-directional differentiation,etc.Cerebral ischemia is caused by neurological impairment,which is the most common cause of death and quality-of-life impairments.The clinical manifestations of the patients with cerebral ischemia are motor function failure,sensory dysfunction and abnormal mental consciousness.A large number of studies have reported that BMSCs transplantation has the therapeutic effects of body sensory and motor function recovery,and can treat ischemic stroke.BMSCs transplantation has brought new hope for the clinical treatment of ischemic cerebrovascular disease.In this paper,the recent progress in the study of BMSCs transplantation for ischemic stroke was reviewed.The mechanism,pathways,influencing factors and clinical application of BMSCs transplantation were summarized.

4.
Basic & Clinical Medicine ; (12): 307-312, 2017.
Article in Chinese | WPRIM | ID: wpr-510536

ABSTRACT

Objective To establish an aging model of rat bone marrow stromal cells (BMSCs) in vitro and in vivo, in order to study the senescence biology of aging BMSCs .Methods The control cell group ( in vitro):isolating, puri-fying and culturing BMSCs from healthy male SD rats .collecting the third generation ( P3) of BMSCs for analysis . The aging model group (in vitro):the P3 BMSCs were incubated with D-Galactose (D-Gal, 30 g/L) for 48 hours. The aging rat model group ( in vivo): the rats were given 120 mg D-Gal by the way of daily neck subcutaneous injection for 42 consecutive days .The control rat group ( in vivo):the rats were administrated with the same volume of saline for the same times .On the second day after the aging model was established , the BMSCs were collecting and culturing for study.1)The proliferative potency was detected by cell counting Kit-8(CCK-8);the distribution of cell cycle and apoptosis was detected by flow cytometry (FCM);2)the ratio of aging BMSCs was examined by the senescence-associated β-Galactosidase(SA-β-Gal) staining;3)malonaldehyde(MDA) content and total super-oxide dismutase(SOD) was examined activity by enzymatic assay; the level of reactive oxygen species (ROS) by DCFH-DA fluorescent staining was counted with FCM;4 ) the expression level of senescence-related signaling was proteins of P16 , P21 , P53 , CDK2 and cyclin D by Western blot .Results Compared with the matched control group, the BMSCs of aging model group displayed a significant decrease in proliferation; the BMSCs were held in G1 phase arrest as the proportion of the cells in G 1 phase increased , while that decreased in S phase ( P<0.05 );and the positive ratio of SA-β-Gal stained BMSCs also significantly increased ( P <0.05 ); BMSCs in the aging model group showed an increasing level of ROS and MDA , meanwhile a decline in total SOD activity was decreased (P<0.05);P16,P21 and P53 protein expression in aging BMSCs was obviously enhanced (P<0.05), at the same time the expression of CDK2 and cyclin D was also decreased ( P<0.05 ) .Conclusions D-Gal can be used to develope an aging model of BMSCs .It acts through up-regulation of expressions of aging-related proteins and in-hibition of oxidative stress injury and chronic inflammation .

5.
Zhonghua zhong liu za zhi ; (12): 885-890, 2017.
Article in Chinese | WPRIM | ID: wpr-809697

ABSTRACT

Objective@#To investigate the influences of bone marrow stromal cells, components of extracellular matrix and cytokine secreted by stromal cells on the chemotherapeutic sensitivity of acute lymphoblastic leukemia cells to cytosine arabinoside (Ara-C).@*Methods@#The co-culture model of acute lymphoblastic leukemia cell Sup-B15 and bone marrow stromal cell OP9 was constructed. Sup-B15 cells were cultured alone or co-cultured with OP9 cells, inactivated OP9 cells, the conditional medium (CM) of co-cultured OP9 cells and Sup-B15 cells, the CM of OP9 cells alone or Sup-B15 cells alone, respectively. The effects of different concentrations of Ara-C on the proliferation of each Sup-B1 cell group mentioned above were detected by cell counting kit-8 (CCK-8) method. The effects of different concentrations of Ara-C on the apoptosis of each group were detected by flow cytometry (FCM). The expressions of Bcl-2 protein in each group were detected by western blot.@*Results@#The results of CCK-8 test showed that the inhibitory efficiency of Ara-C was in a dose-dependent manner. With different concentrations of Ara-C treatment for 48 hours, the half maximal inhibitory concentrations (IC50) of Sup-B15 and OP9 co-cultured group, Sup-B15 and inactivated OP9 co-cultured group were 0.510 and 0.339 μg/ml, respectively, significantly higher than 0.091 μg/ml of Sup-B15 cultured alone group (P<0.05). The IC50 of CM of Sup-B15 and OP9 co-cultured group was 0.204 μg/ml, significantly higher than 0.087 μg/ml of the CM of OP9 cultured alone group (P<0.05) and 0.097 μg/ml of the CM of Sup-B15 cultured alone group (P<0.05). The results of flow cytometry showed that with 0.10 μg/ml Ara-C treatment for 24 hours, the early apoptotic cell percentages of Sup-B15 and OP9 co-cultured group, Sup-B15 and inactivated OP9 co-cultured group and Sup-B15 cultured alone group were (6.67±2.19) %, (8.95±3.04) % and (20.46±2.63) %, respectively. The early apoptotic cell percentages of Sup-B15 and OP9 co-cultured group, Sup-B15 and inactivated OP9 co-cultured group were significantly lower than that of Sup-B15 cultured alone group (P<0.05). The early apoptotic cell percentages of the CM of Sup-B15 and OP9 co-cultured group, the CM of OP9 cultured alone group and the CM of Sup-B15 cultured alone group were (11.16±2.97)%, (22.08±2.71)% and (19.25±1.57)%, respectively, the former two of which were significantly lower than the last one (P<0.05). The results of western blot showed that the relative expression levels of Bcl-2 protein of Sup-B15 cultured alone group, Sup-B15 and OP9 co-cultured group, Sup-B15 and inactivated OP9 co-cultured group, the CM of Sup-B15 and OP9 co-cultured group, the CM of OP9 cultured alone group and the CM of Sup-B15 cultured alone group were 1.00±0.00, 1.53±0.03, 1.38±0.01, 1.26±0.05, 1.03±0.01 and 0.98±0.02, respectively. The expression levels of bcl-2 protein of three combined groups were significantly higher than that of Sup-B15 cultured alone group (P<0.05). while no statistically significant difference was observed between the CM of OP9 cultured alone group and the CM of Sup-B15 cultured alone group (P>0.05).@*Conclusion@#Bone marrow stromal cell OP9, the components of bone marrow extracellular matrix and cytokine secreted by stromal cells are involved in the induction of the chemotherapeutic resistance of Sup-B15 cells to Ara-C.

6.
Article in Chinese | WPRIM | ID: wpr-495321

ABSTRACT

Objective:To investigate the effects of icaritin(ICT)on the proliferation and osteogenic differentiation of rat bone mar-row stromal cells(rBMSCs).Methods:rBMSCs were cultured from the bone marrow of SD rats and identified by multilineage differ-entiation assays.3,6 and 9 days after the treatment of rBMSCs of passage 4 by ICT at 1 0 -9 ,1 0 -8 ,1 0 -7 ,1 0 -6 and 1 0 -5 mol/L re-spectively,the proliferation and differentiation of the cells were examined by cck-8 and alkaline phosphatase (ALP)activity assay kit respectively.The calcium nodule formation was observed by alizarin red(AR)staining 21 days after 1 0 -9 mol/L ICT treatment. Results:Primary rBMSCs showed the typical spindle-like shape with attachment growth.rBMSCs could be induced to osteogenic and adipogenic differentiation.The proliferation of rBMSCs was inhibited but ALP activity was enhanced by ICT.1 0 -9 mol/L ICT in-cresed calcium nodule formation.Conclusion:ICT can dose-dependently inhibit the proliferation,but promote the osteogenic differ-entiation of rBMSCs.

7.
Article in English | WPRIM | ID: wpr-651482

ABSTRACT

In this study, we developed the disc-type bio-cartilage reconstruction strategies for transplantable hyaline cartilage for reconstructive surgery using 3D-cell sheet culture of human bone marrow stromal cells and human costal chondrocytes. We compared chondrogenesis efficiency between different chondrogenic-induction methods such as micromass culture, pellet culture, and 3D-cell sheet culture. Among them, the 3D-cell sheet culture resulted in the best chondrogenesis with the disc-type bio-cartilage (>12 mm diameter in size) in vitro, but sometimes spontaneous curling and contraction of 3D-cell sheet culture resulted in the formation of bead-type cartilage, which was prevented by type I collagen coating or by culturing on amniotic membrane. Previously, it was reported that tissue-engineered cartilage reconstructed in vitro does not maintain its cartilage phenotype after transplantation but tends to transform to other tissue type such as bone or connective tissue. However, the disc-type bio-cartilage of 3D-cell sheet culture maintained its hyaline cartilage phenotype even after exposure to the osteogenic-induction condition in vitro for 3 weeks or after the transplantation for 4 weeks in mouse subcutaneous. Collectively, the disc-type bio-cartilage with 12 mm diameter can be reproducibly reconstructed by the 3D-cell sheet culture, whose hyaline cartilage phenotype and shape can be maintained under the osteogenic-induction condition as well as after the transplantation. This disc-type bio-cartilage can be proposed for the application to reconstructive surgery and repair of disc-type cartilage such as mandibular cartilage and digits.


Subject(s)
Animals , Humans , Mice , Amnion , Bone Marrow , Cartilage , Chondrocytes , Chondrogenesis , Collagen Type I , Connective Tissue , Hyalin , Hyaline Cartilage , In Vitro Techniques , Mesenchymal Stem Cells , Phenotype
8.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;46(1): 39-51, 11/jan. 2013. tab, graf
Article in English | LILACS | ID: lil-665801

ABSTRACT

Imatinib mesylate (IM) is used to treat chronic myeloid leukemia (CML) because it selectively inhibits tyrosine kinase, which is a hallmark of CML oncogenesis. Recent studies have shown that IM inhibits the growth of several non-malignant hematopoietic and fibroblast cells from bone marrow (BM). The aim of the present study was to evaluate the effects of IM on stromal and hematopoietic progenitor cells, specifically in the colony-forming units of granulocyte/macrophage (CFU-GM), using BM cultures from 108 1.5- to 2-month-old healthy Swiss mice. The results showed that low concentrations of IM (1.25 µM) reduced the growth of CFU-GM in clonogenic assays. In culture assays with stromal cells, fibroblast proliferation and α-SMA expression by immunocytochemistry analysis were also reduced in a concentration-dependent manner, with a survival rate of approximately 50% with a dose of 2.5 µM. Cell viability and morphology were analyzed using MTT and staining with acrydine orange/ethidium bromide. Most cells were found to be viable after treatment with 5 µM IM, although there was gradual growth inhibition of fibroblastic cells while the number of round cells (macrophage-like cells) increased. At higher concentrations (15 µM), the majority of cells were apoptotic and cell growth ceased completely. Oil red staining revealed the presence of adipocytes only in untreated cells (control). Cell cycle analysis of stromal cells by flow cytometry showed a blockade at the G0/G1 phases in groups treated with 5-15 µM. These results suggest that IM differentially inhibits the survival of different types of BM cells since toxic effects were achieved.


Subject(s)
Animals , Male , Mice , Antineoplastic Agents/pharmacology , Benzamides/pharmacology , Hematopoietic Stem Cells/drug effects , Mesenchymal Stem Cells/drug effects , Piperazines/pharmacology , Pyrimidines/pharmacology , Cell Proliferation , Cell Survival , Cells, Cultured , Colony-Forming Units Assay , Fibroblasts , Hematopoietic Stem Cells/cytology , Immunohistochemistry , Mesenchymal Stem Cells/cytology
9.
Chinese Journal of Neuromedicine ; (12): 775-779, 2012.
Article in Chinese | WPRIM | ID: wpr-1033591

ABSTRACT

Objective To observe the expression of growth factors (vascular endothelial growth factor [VEGF],stromal cell-derived factor-1 [SDF-1 ],basic fibroblast growth factor [bFGF],insulin-like growth factor [IGF-1],transforming growth factor-β [TGF-β],platelet-derived growth factor [PDGF],brain derived neurotrophic factor [BDNF],glial cell line-derived neurotrophic factor [GDNF] and nerve growth factor [NGF]) in rat ischemic brain tissues after intravenous implantation of bone marrow stromal cells (BMSCs) and/or endothelial progenitor cells (EPCs). Methods Healthy adult Wistar rats were randomly divided into 4 groups:vehicle group,BMSCs transplantation group,EPCs transplantation group and BMSCs combined with EPCs transplantation group (n=20). The rats were subjected to middle cerebral artery occlusion (MCAO),and 24 h after that,they were intravenously transplanted with either 3×106 BMSCs,EPCs,BMSCs/EPCs or 1 mL physiological saline.Seven d after transplantation,real time-PCR and Western blotting were employed to detect the expressions of VEGF,SDF-1,bFGF,IGF-1,TGF-β,PDGF-BB,BDNF,GDNF and NGF. Results The mRNA expressions of bFGF,VEGF and BNDF in the BMSCs/EPCs transplantation group were significantly higher as compared with those in the other groups (P<0.05).BMSCs transplantation group enjoyed the highest mRNA levels of NGF,GDNF and TGF-β among all the groups, significantly higher as compared with those in the other groups (P<0.05),followed by BMSCs/EPCs transplantation group.EPCs transplantation group enjoyed the highest mRNA levels of PDGF,IGF-1 and SDF-1,significantly higher as compared with those in the other groups (P< 0.05), followed by BMSCs/EPCs transplantation group. Conclusion BMSCs combined with EPCs implantation can promote the functional rehabilitation in rats after focal cerebral ischemia, which provides new way for improving the transplantation success rate.

10.
Article in Chinese | WPRIM | ID: wpr-1033170

ABSTRACT

Objective To evaluate whether bone marrow stromal cells (BMSCs) transplantation can improve the cognitive function of aged rats with vascular dementia. Methods Thirty rats were equally ramdomized into normal control group and 4 treatment groups; the 4 treatment groups received subcutaneous injection of D-galactose (D-gal) for 4 weeks; and then, two-vessel occlusion (2VO) was performed in 3 of the treatment groups; and 24 h after 2VO, D-gal+2VO+saline group and D-gal+2VO+BMSCs group were subjected to stereotactic injection of normal saline and BMSCs into the subventricular zone (SVZ), respectively. The cognitive function was examined by Morris water maze test 6 weeks after stereotactic injection; immunofluorescence staining was employed to observe the transplantation ratio of BMSCs to neurons. Results Increased times and distances during Morris water maze in rats of the D-gal+2VO group, D-gal+2VO+saline group and D-gal+2VO+BMSCs group were noted as compared with those in the controls, indicating that the cognitive function of rats in these 3 groups was obviously impaired; these rats had the characteristics of having vascular dementia.Transplanted BMSCs in the D-gal+2VO+BMSCs group distributed around the lateral ventricles, and acquired the phenotypes of neurons (2%) and astrocyte (1%) 6 weeks after the transplantation. In addition, compared with that in rats of the D-gal+2VO group and D-gal+2VO+saline group, the cognitive dysfunction of rats in the D-gal+2VO+BMSCs group was improved (needing less time and swimming shorter distance, no difference in speed of swimming). Conclusion The D-gal injection plus 2VO can result in cognitive dysfunction of rats, and the engrafted BMSCs may exhibit the beneficial effect on cognitive function.Neural function remolding caused by interaction between BMSCs and host brain may be responsible for the function improvement

11.
Academic Journal of Xi&#39 ; an Jiaotong University;(4): 25-29, 2010.
Article in Chinese | WPRIM | ID: wpr-844746

ABSTRACT

Objective: To investigate the effects of Panax notoginseng saponins (PNS) on hydrogen peroxide (H2O2)-induced apoptosis in cultured rabbit bone marrow stromal cells (BMSCs). Methods: BMSCs from 3-month-old New Zealand rabbits were isolated and cultured by the density gradient centrifugation combined with adherent method. The cultured BMSCs were divided into three groups: normal control, H2O2 treatment (100 μmol/L), and PNS pretreatment (0.1 g/L). Intracellular reactive oxygen species (ROS) levels as the index of oxidative stress were measured by using 2′7′-dichlorodihydrofluorescein diacetate. Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/PI. The protein expression of Bax in BMSCs was analyzed by Western blotting. Activity of caspase-3 enzyme was measured by spectrofluorometry. Results: Pretreatment with PNS significantly decreased intracellular ROS level induced by H 2O2 (P<0.01). PNS markedly attenuated H 2O2-induced apoptosis rate from 38.68% to 19.24% (P<0.01). PNS reversed H2O2-induced augmentation of Bax expression. Furthermore, PNS markedly reduced the altered in activity of caspase-3 enzyme induced by H2O2 (P<0.01). Conclusion: PNS has a protective effect on hydrogen peroxide-induced apoptosis in cultured rabbit BMSCs by scavenging ROS and decreasing Bax expression and caspase-3 activity.

12.
Article in Chinese | WPRIM | ID: wpr-621631

ABSTRACT

Objective To investigate the effects of Panax notoginseng saponins (PNS) on hydrogen peroxide (H2O2)-induced apoptosis in cultured rabbit bone marrow stromal cells (BMSCs). Methods BMSCs from 3-month-old New Zealand rabbits were isolated and cultured by the density gradient centrifugation combined with adherent method. The cultured BMSCs were divided into three groups: normal control, H2O2 treatment (100μmol/L), and PNS pretreatment (0.1g/L). Intracellular reactive oxygen species (ROS) levels as the index of oxidative stress were measured by using 2'7'-dichlorodihydrofluorescein diacetate. Flow cytometry was used to observe the apoptosis of BMSCs by staining with annexinV-FITC/PI. The protein expression of Bax in BMSCs was analyzed by Western blotting. Activity of caspase-3 enzyme was measured by spectrofluorometry. Results Pretreatment with PNS significantly decreased intracellular ROS level induced by H2O2 (P<0.01). PNS markedly attenuated H2O2-induced apoptosis rate from 38.68% to 19.24%(P<0.01). PNS reversed H2O2-induced augmentation of Bax expression. Furthermore, PNS markedly reduced the altered in activity of caspase-3 enzyme induced by H2O2(P<0.01). Conclusion PNS has a protective effect on hydrogen peroxide-induced apoptosis in cultured rabbit BMSCs by scavenging ROS and decreasing Bax expression and caspase-3 activity.

13.
Article in Korean | WPRIM | ID: wpr-197402

ABSTRACT

PURPOSE: Major drawbacks of conventional bone marrow stromal cells (BSCs) transplantation method are mainly caused by direct transplanted cell to host cell interactions. We hypothesized that separation of the transplanted cells by a microporous membrane might inhibit most of the potential adverse effects and induce superior effect. The purpose of the study is to determine the optimal condition of the microporous membrane. METHODS: First, BSCs were placed in polyethylene terephthalate (PET) transwell inserts with 3, 8, or 12 micrometer pore size, and cultured in 24 well culture plates. After 5 days, bottoms of the plates were observed for presence of attached BSCs in monolayer and cell numbers were evaluated. Second, BSCs were placed PET, polycarbonate (PCT), and mixed cellulose esters (MCE) transwell inserts with 3 and 8 micrometer pore size, and cultured in 24 well culture plates. After 3 days, the supernatants of the media left in culture plate were analyzed for collagen, vascular endothelial growth factor (VEGF), platelet derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF). Third, BSCs were placed in 15% and 70% of the PET membrane with 3 micrometer pore size. All the experimental conditions and methods were same as the second study. RESULTS: The optimal pore sizes to prevent BSC leakage were 3 micrometer and 8 micrometer. The amounts of type I collagen and three growth factors tested did not show significant differences among PET, PCT, and MCE groups. However, the collagen, VEGF, and bFGF levels were much higher in the high (70%) density group than in the low (15%) density group. CONCLUSION: This study revealed that the optimal pore size of membrane to prevent direct BSC to recipient cell contact is in between 3 micrometer and 8 micrometer. Membrane materials and pore sizes do not influence the collagen and growth factor passage through the membrane. The most striking factor for collagen and growth factor transport is pore density of the membrane.


Subject(s)
Bone Marrow , Cell Communication , Cell Count , Cellulose , Collagen , Collagen Type I , Esters , Fibroblast Growth Factor 2 , Intercellular Signaling Peptides and Proteins , Membranes , Mesenchymal Stem Cells , Phthalic Acids , Platelet-Derived Growth Factor , Polycarboxylate Cement , Polyethylene Terephthalates , Strikes, Employee , Transplants , Vascular Endothelial Growth Factor A , Wound Healing
14.
Article in Chinese | WPRIM | ID: wpr-395767

ABSTRACT

Objective To investigate the efficacy of the combination of bone marrow stromal cells (BMSCs) and bcl-2 gene in the treatment cerebral ischemia taxi its effect on the expression of basic fibroblast growth factor (bFGF) in rats.Methods Forty Wistar rats were used to establish middle cerebral artery occlusion model. They were randomly divided into 4 groups: saline control, bcl-2, BMSCs and BMSCs + bcl-2 groups (n = 10 in each group). Every group was redivided into 3- and 14-day after reperfusion subgroups (n = 5 in each subgroup). Neurological scores of the experimental rats were assessed. BMSCs were labeled with bromodeoxyuridine (BrdU). The distribution and numbers of BMSCs, the expressions of Bcl-2 and bFGF were detected by immunohistochemistry, and apoptotic cells were detected with TUNEL staining in rat brain. Results The neurological score at day 3 after reperfusion in the BMSCs + bcl-2 group was significantly lower than that in the saline control group (P < 0. 05), and at day 14, it was significantly lower than that in the other 3 groups (all P <0. 05). A large number of BrdU-positive BMSCs were observed in the infarcted hemisphere in the BMSCs + bcl-2 and BMSCs groups. The numbers of BrdU-positive BMSCs at day 3 and 14 after reperfusion in the BMSCs + bcl-2 group were significantly higher than those in the BMSCs group (all P <0. 05). The expressions of Bcl-2 in the infarcted hemisphere at day 3 and 14 after reperfusion in BMSCs +hel-2 group wre significantly higher than those in the other 3 groups (all P <0. 05). The expressions of Bcl-2 at all time points were increased more significantly than those in the other 3 groups (all P <0. 05). The numbers of apoptosis in brain at all time points in the BMSCs + bcl-2 group were decreased more significantly than those in the other 3 groups (all P <0. 05). Conclusions Both BMSCs and bcl-2 genes have the therapeutic effect on cerebral ischemia. The efficacy of combination of both ,of them is significantly superior to monotherapy. They may significantly improve the neurological function and increase the expression of bFGF in rats. Its mechanism may be that bcl-2 genes have inhibited BMSCs apoptosis at the same time of anti-apoptosis in brain.

15.
Tumor ; (12): 1081-1085, 2008.
Article in Chinese | WPRIM | ID: wpr-849247

ABSTRACT

Objective: To investigate the effects of bone marrow stromal cells (BMSCs) isolated from leukemia patients on the invasion capacity leukemia SHI-1 cells in vitro and the underlying mechanism. Methods: BMSCs were isolated from leukemia patients and their conditional culture medium were collected. The expressions of extracellular matrix metalloproteinase inducer (EMMPRIN) in SHI-1 cells and BMSC were detected by RT-PCR. The BMSC and SHI-1 cells were mixed at 1: 10 and inoculated in Matrigel-coated transwells. Then CXC chemokine receptor 4 (CXCR4, final concentration of 2 μg/mL) or functional antibody of EMMPRIN were added. The BMSC cultured medium was used as control on cell invasion test. The alteration of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase 2 (TIMP-2), and CXCR4 mRNA were determined by real-time PCR before and after co-culture of SHI-1 cells and BMSC. The content of stromal cell-derived factor 1 (SDF-1) in serum-free supernatent was measured by ELISA. Results: Both SHI-1 and BMSC expressed EMMPRIN. The invasion capacity of SHI-1 cells increased significantly after co-culture with BMSC, which could be blocked by CXCR4 and functional antibody of EMMPRIN. The cultured medium of BMSC did not increase the invasion capacity of SHI-1 cells. The mRNA expression levels of MMP-2, MMP-9, TIMP-2, and CXCR4 as well as SDF-1 contents in serum-free supernatent increased significantly after the SHI-1 cells were co-cultured with BMSC. Conclusion: When BMSC islated from leukemia patients contacted with leukemia SHI-1 cells, they increases the invasion capacity of SHI-1 cells through multiple molecule pathways on the surface of them on cell invasion test. It may be an important mechanism responsible for the invasion of leukemia cells to the outer space of bone marrow.

16.
Article in Chinese | WPRIM | ID: wpr-401911

ABSTRACT

Gene therapy refers to the introduction of normal genes into human target cells for correcting gene defects or exerting therapeutic action,and thus achieves the goal of treatment of disease.Bone marrow stromal cells (BMSCs) are stem cells that possess self-renewal and multi-directional differentiation potential and easy to amplify in vitro,and they also express many therapeutic exogenous genes in vitro or in vivo.So BMSCs have been regarded as an ideal target cell of cell and gene therapy.This article reviews the biological characteristic of BMSCs,some commonly used gene therapy vectors and their applications in gene therapy of central nervous system diseases.

17.
Article in Korean | WPRIM | ID: wpr-113022

ABSTRACT

PURPOSE: The aim of this study is to compare the effects of bone marrow stromal cells(BSCs) and fibroblasts on wound healing activity in vivo, especially on epithelization. METHODS: The fibroblasts and BSCs were harvested from patients and cultured. Ten Spague-Dawley white rats were used. A 5 mm punches were made to excise skin and subcutaneous tissue in a round fashion at six sites on the back area of each rat. Four hundred thousand cells suspended in 0.05 ml fibrinogen were applied to the created wounds. The cells in group I, II, and III were no cells, fibroblasts and BSCs. The lengths of epithelial gap at the widest wound site were compared with autopsy specimens obtained on the 6th day after cell therapy under light microscope. Statistical comparisons were performed using the Mann-Whitney U-test, and the p value<0.05 was considered statistically significant. RESULTS: The best epithelization was also seen in the BSC group, followed by fibroblast and no cell groups. CONCLUSION: These results demonstrate that BSC has superior effect on stimulating wound healing than fibroblast, which is currently used for wound healing.


Subject(s)
Animals , Humans , Rats , Autopsy , Bone Marrow , Cell- and Tissue-Based Therapy , Fibrinogen , Fibroblasts , Mesenchymal Stem Cells , Skin , Subcutaneous Tissue , Wound Healing , Wounds and Injuries
18.
Article in Korean | WPRIM | ID: wpr-24495

ABSTRACT

PURPOSE: In the previous in vitro studies the bone marrow stromal cells(BSCs) have shown the superior effect for wound healing activity than fibroblasts, which includes cell proliferation, type I collagen synthesis, and the production of bFGF, VEGF and TGF-beta in chronic wound healing. The aim of this study is to compare the effects of BSCs and fibroblasts on wound healing activity in vivo, especially on collagen synthesis. METHODS: The fibroblasts and BSCs were harvested from patients and cultured. The cultured cells were infiltrated into the pores of polyethylene discs. These discs were divided into three groups according to the mixed cells. In groups I, II and III the discs were loaded with no cells, fibroblasts and BSCs, respectively. Twelve discs per group(total 36 discs) were made for this study. After creating 6 pockets in the back of each rats, each discs was implanted into each pockets. At three time intervals from 1 to 3 weeks, the implanted discs were harvested for the histological and quantitative analysis. The amount of collagen produced was evaluated using ELISA. Statistical comparisons were made using the Mann-Whitney U-test. RESULTS: There was great difference in the collagen synthesis among the three groups by the 1st and 2nd weeks. The BSC group showed highest collagen level, followed by fibroblast group and no cell group(p<0.05). The 3rd week specimens also showed greater collagen amount in BSC and fibroblast groups compared to those of no cell group(p<0.05). However, there was little difference between BSC and fibroblast groups. CONCLUSION: This result demonstrates that BSC has superior effect on stimulating wound healing than fibroblast, which is currently used for wound healing.


Subject(s)
Animals , Humans , Rats , Bone Marrow , Cell Proliferation , Cells, Cultured , Collagen Type I , Collagen , Enzyme-Linked Immunosorbent Assay , Fibroblasts , Mesenchymal Stem Cells , Polyethylene , Transforming Growth Factor beta , Vascular Endothelial Growth Factor A , Wound Healing
19.
Article in Chinese | WPRIM | ID: wpr-543979

ABSTRACT

[Objective]To evaluate the effect of the tissue engineered nerve for bridging and repairing verve defect.[Method]Human bone marrow stromal cells(hBMSCs) were purified with centrifugate method,cultured in DMEM,induced with ATRA,BDGF and affected by heregulin,forsholin,bFGF(basic fibroblast growth factor),and PDGF(platelet-derived growth factor) of hBMSCs).The protein positive rate of S100 and GFAP of hBMSCs were determined by immunohistochmical staining.The tissue engineered nerves were constructed with hBMSCs mixed with extra-cellular matrix(ECM) and polylactic acid(PLA) tube.A 10mm defect of sciatic nerve was created in 24 Wistar mouse right limbs and ramdonly divided into three groups: group A(n=8),nerve defects bridged with polylactic acid(PLA) tube containing induced Schwann cells mixed with ECM,group B(n=8),with PLA tube containing ECM,group C(n=8) with autologous nerve graft.Functional recovery of nerve was examined by electrophysiological method and histological changes were examined with histological stainning of nerve and measurement quantity of new axon.[Result]The Schwann cells were presented at 12 wks after operation.The histologic and functional recovery of nerves of group A and group C were better than those of group B.the showed significant difference between group A or group C and group B and no significant difference between group A and group C.(PAB=0.021,PBC=0.001,PAC=0.065).Degradation of PLA tubes showed in group A and group B.[Conclusion]Schwann cells could be induced from hBMSCs,and the tissue engineered nerves,which were contructed by induced Schwann cells mixed with ECM and PLA tube,could be used to bridged and repair the peripheral nerve defect.

20.
Article in Chinese | WPRIM | ID: wpr-544426

ABSTRACT

[Objective]To investigate the repairing effect and mechanism for treatment of femoral head necrosis on transplantation of bone marrow stromal cells.[Method]Twenty-four Newzland rabbits were divided into two groups randomly,Group A as core decompression group and Group B as Bone marrow stromal cells autograft group.Animal femoral head neorosis model was made by freezing method with liquid nitrogen.After freezing and drilling,The drilled necrotic cavities of Group A was implanted with gelatin sponge,while of Group B was with gelatin sponge combined by bone marrow stromal cells.Three animals in every group were sacrificed respectively at 2,4,6,8 weeks and subjected to radiograph and microscope examination.[Result](1)Radiograph examination:At 2 weeks,the drilled holes in Group A and B all presented low density images.At 4 weeks,the density in the drilled holes increased in group B and the density around the drilled holes increased in group A.At 8 weeks,sclerotic line formed around the drilled holes in Group A and trabeculae appeared in the drilled holes in group B.(2)Microscope examination:in group A,at 2 weeks,a few inflamatory cells appeared in the holes with many osteoblasts and ostoid appeared around the drilled holes.At 8 weeks,marrow formed in the drilled holes in group B,at 2 weeks,there were a large amount of osteoblasts in the drilled holes and some ostoid formed around thc drilled holes.At 4 weeks,the drilled holes were full of trabeculae.At 8 weeks,the trabeculae matured with the marrow appeared in the intertrabecular space.[Conclusion]Bone marrow stromal cells autografting may be an effective way to treat rabbit femoral head necrosis.

SELECTION OF CITATIONS
SEARCH DETAIL