Research Advances on the Role of Bone Marrow Stromal Cell in Acute Lymphoblastic Leukemia --Review / 中国实验血液学杂志

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.

Senescence induced by D-galactose and its biological mechanism in rat bone marrow stromal cells / 基础医学与临床

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 .

Bone marrow stromal cells as a therapeutic treatment for cerebral ischemic / 国际生物医学工程杂志

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.

Effects of bone marrow stromal cells on the chemotherapeutic sensitivity of acute lymphoblastic leukemia cells / 中华肿瘤杂志

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.

The effects of icaritin on the proliferation and osteogenic differentiation of rat bone marrow stromal cells / 实用口腔医学杂志

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.

Disc-Type Hyaline Cartilage Reconstruction Using 3D-Cell Sheet Culture of Human Bone Marrow Stromal Cells and Human Costal Chondrocytes and Maintenance of Its Shape and Phenotype after Transplantation

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.

Construction of Nfic gene 3′UTR dual luciferase reporter vector and targeting verification between Nfic and miR-20a / 天津医药

Objective To construct a luciferase reporter vector containing the 3′untranslated region (3′UTR) of nuclear factor I-C (nuclear factor I-C, Nfic), and apply dual luciferase reporter gene system to determine the association between microRNA-20a (miR-20a) and its potential target gene Nfic. Methods The potential complementary binding sites of miR-20a and Nfic were predicted by Targetscan. The 3′UTR of Nfic fragment amplified by PCR was cloned into luciferase reporter vector MIR- Report Luciferase. The luciferase reporters containing 3′ UTR of Nfic and miR- 20 mimics (experimental group) or NC mimics (control group) were co-transfected into 293-AD cells. Cells were collected, and then dual-luciferase reporter assay was performed to detect the luciferase activity of the two groups of cells, consequently the relationship between miR-20a and Nfic was identified. The miR-20a mimics and NC mimics were transfected into marrow stromal cell line ST2 respectively. The total cell lysates were collected, and the expression level of NFIC was detected by Western blotting assay. Results Results of double enzyme digestion and DNA sequencing showed that sequence of luciferase reporter vector was correct. miR-20a specificity bounded to Nfic 3′UTR and inhibited the luciferase activity of the reporter construct (P<0.05). Western blotting assay showed that the NFIC protein level was obviously down-regulated in ST2 cells after the transfection of miR-20a mimics compared with that of control. Conclusion The luciferase reporter vector containing the 3′UTR of Nfic is constructed successfully, which confirms that miR-20a can direct effect on Nfic3′UTR and repress its luciferase activity.

In vitro inhibitory effects of imatinib mesylate on stromal cells and hematopoietic progenitors from bone marrow

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.

CXCR4 expression of bone marrow CD34+cells in myelodysplastic syndromes and its correlation with cell migration / 中国肿瘤临床

Objective:To evaluate the expression of CXCR4 and the migration rate of bone marrow stromal CD34+cells in differ-ent risk groups with myelodysplastic syndromes (MDS) using correlation analysis. Methods: Forty MDS patients were divided into low-and high-risk groups based on the International Prognosis Scoring System (IPSS). The former was composed of 20 patients with IPSS0.05). The migration rate of CD34+cells on the ef-fects of SDF-1αand marrow stromal cells were significantly increased in the high-risk MDS group compared with those in the low-risk and control groups (P<0.000 1). Migration rate of CD34+cells on the effect of marrow stromal cells was positively correlated with CX-CR4 expression (P=0.000 1). Conclusion:The CXCR4 expression and migration rates of CD34+cells on the effect of marrow stromal cells are significantly higher in the high-risk MDS group than in the low-risk group. Migration rate has a positive correlation with the CXCR4 expression, which further indicates that MDS is a heterogeneous group of hematopoietic stem cell malignancies. The expres-sion and function of SDF-1 and its receptor CXCR4 differ within each group with various risks. SDF-1 and CXCR4 may be involved in MDS pathogenesis.

Effects of Panax notoginseng saponins on hydrogen peroxide-induced apoptosis in cultured rabbit bone marrow stromal cells / 药物分析学报

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.

Optimal Condition of Microporous Membrane for Bone Marrow Stromal Cell Allotransplantation to Stimulate Wound Healing in Vitro

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.

Effects of Panax notoginseng saponins on hydrogen peroxide-induced apoptosis in cultured rabbit bone marrow stromal cells / 西安交通大学学报·英文版

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.

Effect of combination of bone marrow stromal cells and bcl-2 gene therapy on expression of basic fibroblast growth factor in focal cerebra ischemic rat brain / 国际脑血管病杂志

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.

Granulocyte Colony Stimulating Factor for Focal Cerebral Ischemic Injury / 国际脑血管病杂志

Granulocyte colony stimulating factor(G-CSF)can mobilize hemopoietic stem cells(HSCs)from bone marrow to peripheral circulation,stimulating HSCs and marrow stromal cells(MSCs)homing to the lesion of cerebral ischemia,contributing to the interactions of HSCs and MSCs with the cells in ischemic penumbra,and promoting the generation of neurotrophic factors.G-CSF also has antiapoptotic effect on neurons and drives neurogenesis.Mobilizing autologous HSCs and MSCs using G-CSF for the treatment of local cerebral ischemic injury may reduce the volume of cerebral infarction,and improve neurological functional recovery.

Applications of Bone Marrow Stromal Cells in Gene Therapy of Central Nervous System Diseases / 国际脑血管病杂志

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.

Bone marrow stromal cells isolated from leukemia patients promotes the invasion capacity of SHI-1 cells in vitro and its underlying mechanism / 肿瘤

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.

Effect of Transplantation of Human Bone Marrow Stromal Cells or Dermal Fibroblasts on Wound Healing

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.

Effect of Transplantation of Bone Marrow Stromal Cells and Dermal Fibroblasts on Collagen Synthesis

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.

Expression of VEGF gene mediated by adenovirus in human bone marrow stromal cell / 中国矫形外科杂志

[Objective]To investigate the possibility of stable expression of VEGF~65 in human bone marrow stromal cell.[Method]hBMSc were divided into transfection group, empty vector group and control group, hBMSc were infected with Ad-VEGF165(MOI=20 pfu). The hBMSc proliferation and transfection efficiency were confirmed by fluorescence microscope.The expression of VEGF165 gene of cultured hBMSc transfected with VEGF165 gene was assayed by RT-PCR, ELISA and Western blot.[Result]VEGF165 gene was amplified with RT-PCR.GAPDH gene(internal control) was located at 549 bp in the three groups, VEGF165 gene was located at 500 bp in transfection group but not in the other groups. ELISA assay showed that the expression had significant difference in the transfection group compared with the empty vector group and control group (P

Experimental study on treatment of femoral head necrosis in rabbit with transplantation of bone marrow stromal cells / 中国矫形外科杂志

[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.