Objective To review research progress of adipose tissuederived stromal cells (ADSCs).Methods The recent articles on ADSCs were extensively reviewed, and the culture and differentiation ability of ADSCs were investigated.Results A population of stem cells could be isolated from adult adipose tissue, they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling. The majority of the isolated cells were mesenchymal origin, with a few pericytes,endothelial cells and smooth muscle cells. ADSCs could be induced to differentiate intomultiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic and adipogenic cells, they could also differentiate into nerve cells.Conclusion ADSCs can substitute mesenchymal stem cells and become an alternative stem cells source for tissue engineering.
Objective To investigate the expression of stromal cell derived factor-1 ( SDF-1) and the effects of budesonide suspension for inhalation ( Pulmicort Respules) in mice with asthma. Methods Thirty Kunming female mice were randomly divided into three groups, ie. a control group, an asthma group, and a pulmicort treatment group. The asthma group and the pulmicort treatment group were sensitized with ovalbumin ( OVA) by a combination of intraperitoneal injection and repeated OVA intranasal challenges to establish mouse asthma model. The pulmicort treatment group received 100μL pulmicort by intranasal administration before OVA challenge. The immunohistochemistry was used to estimate the expression of SDF-1 in lung tissues. HE staining and Wright-Giemsa staining method were used to assess inflammatory infiltration in the airway and bronchoalveolar lavage fluid ( BALF) respectively. Results The expression of SDF-1 in the asthma group increased significantly compared with the control group ( 0.48 ±0.03 vs. 0.21 ± 0.02, Plt;0.05) , and significantly decreased after the intervention with pulmicort ( 0.29 ±0.01 vs. 0.48 ± 0.03, Plt; 0.05 ) . Compared with control group, the infiltration of inflammatory cells in airway was significantly enhanced in the asthma group, and attenuated in the pulmicort treatment group. The total number of inflammatory cells and eosinophil, lymphocyte, neutrophil counts in BALF increased significantly in the asthma group compared with the control group, and decreased significantly after pulmicort intervention. Conclusion SDF-1 may play an important role in the recruitment of inflammatory cells in asthmatic airway and pulmicort may relieve airway inflammation by decreasing the expression of SDF-1.
【Abstract】Objective Stromal cell-derived factor-1(SDF-1, CXCL12) is a member of the CXC subfamily of chemokines which, through its cognate receptor (CXCR4), plays an important role in tumor invasion and metastasis. This study analyzed quantitatively the expression of SDF-1 and its relation with clinicopathologic feature and clinical outcome in human breast cancer.Methods Expression of SDF-1 mRNA in 8 breast cancer cell lines, an endothelial cell line HECV and a fibroblast cell MRC5 was studied by using RT-PCR. In addition, the expression of SDF-1 was investigated at both protein (immunohistochemistry) and mRNA(real-time PCR) levels in a group of human normal mammary(n=32) and tumour tissues(n=120). Results SDF-1 expression was identified in MRC5, MDA-MB435s, MDA-MB436, MCF7 cell lines, breast tumour and normal tissues. Significantly higher level of SDF-1 was seen in lymph node positive than in lymph node negative tumours (399.00±210.00 vs 0.89±0.47), P=0.048. The level of SDF-1 expression in patients who developed local recurrence or metastasis, or patients who died of breast cancer was higher than in patients who were disease free as well, (670.00±346.00 vs 0.83±0.35), P=0.01. It was most notable that level of SDF-1 was significantly correlated with over survival (P=0.01) and incidence free survival (P=0.035, by Cox proportion analysis).Conclusion SDF-1 is a factor that is expressed in both stromal cells and some breast cancer cells. Its level are correlated with lymph node involvement, prognosis and survival in patients with breast cancer. SDF-1 may therefore have a potential prognostic value in breast cancer.
Objective To investigate the surgical treatment effect for patients with gastrointestinal stromal tumor (GIST) of the rectum and its clinical characteristics. Methods The medical records of 22 patients who had undergone surgery for GIST of the rectum between March 2003 and February 2010 in this hospital were analyzed. Results There were 14 males and 8 females with a median age of 51 years (range 27－81 years). There were 12 patients without symptoms, 10 patients with clinical symptoms, included: hematochezia 4 cases, difficult defecation 2 cases, shape of defecate change 2 cases, crissum pain 1 case, times of defecate increase 1 case. Course of disease was 2 weeks-18 months with average 6 months. All patients underwent curative resection: in form of abdominoperineal resection in 3 patients, transanal excision in 8 patients, Mason operation in 8 patients, and transanal endoscopic microsurgery in 3 patients. The median tumor size was 3.1 cm (range 0.4－18.5 cm). The diameter of tumor lt;2.0 cm was 11 cases, 2.1-5.0 cm was 8 cases, 5.1-10.0 cm was 2 cases, gt;10.0 cm was 1 case. Twentyone of 22 cases were positive for CD117, 18 cases positive for CD34, 5 cases positive for αsmooth muscle actin (SMA), and 2 cases positive for Desmin. Local recurrence or hepatic metastasis developed in 2 patients with average 26 months of follow-up (range 1 month to 7 years), and who were then treated with imatinib for more than 1 year. Conclusions The primarily treatment of rectal GIST is surgical. Imatinib therapy is effective against local and systemic recurrent GIST of the rectum.
ObjectiveTo explore the methods of separation， culture， and identification of breast cancer stromal fibroblasts (BCSFs), which could build up a good basis for the further research of function. MethodsBreast cancer tissues were obtained during breast cancer operation， and were cut into pieces with size of 1 mm×1 mm×1 mm under aseptic conditions， then the pieces of the tissues were digested by collagenase Ⅰ and hyaluronidase. Finally the cells separated from the tissues incubated at 37 ℃ with 5% CO2 and 95% air humidified incubator. Morphological characteristics of the fibroblasts were observed under light microscope. The certain proteins were examined by immunohistochemistry （using CK， Vimentin， α-SMA, and TE-7 antibody） and flow cytometric analysis (CD34 and CD45). ResultsThe separated cells begin to attach to the wall of flask within 24 h and reached almost confluency in about 7 d to 10 d . According to identification, the successful rate of separation and culture of BCSFs was 90%（18/20）, and the characteristics of cells showed that morphological characteristics of the fibroblasts was flat spindle， rich cytoplasm, and a flat ovoid cystic nuclear. The fibroblasts in breast cancer tissues showed negative staining for cytokeratin， positive staining for vimentin， alpha-smooth muscle actin, and TE-7， and negative for CD34 and CD45 by flow cytometric analysis. ConclusionsThe fibroblasts in breast cancer tissues could be easily obtained by tissues cuting combined enzyme digestion and rocking technology in vitro. The present study provide an experimental foundation for further studies on fibroblasts in breast cancer.
ObjectiveTo investigate the relationship between the level of stromal cell-derived factor-1 (SDF-1), internal carotid artery stiffness index, and non-arteritic anterior ischemic optic neuropathy (NAION) with macular edema (ME). MethodsA retrospective study. A total of 202 patients with NAION diagnosed by ophthalmic examination in Department of Ophthalmology, The Second Affiliated Hospital of Jiamusi University from January 2023 to January 2025 were included in the study. Based on the presence or absence of ME, the patients were divided into the NAION+ME group and the NAION group, with 94 and 108 cases respectively. A prediction model was constructed based on the influencing factors. To comprehensively evaluate the predictive value of SDF-1 level and carotid artery stiffness index for NAION with ME, a multidimensional analytical approach was employed. The diagnostic performance of individual and combined markers was assessed by constructing receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC). Multivariate logistic regression analysis was performed to determine their independent predictive value. Stratified subgroup analyses were conducted to explore predictive differences across various populations. Cox proportional hazards regression models were established to evaluate long-term predictive value. Restricted cubic spline (RCS) analysis was applied to reveal potential nonlinear dose-response relationships. Mediation effect models were constructed to analyze the mediating role of carotid artery stiffness index in the association between SDF-1 level and NAION with ME. ResultsIn the NAION+ME group, systolic blood pressure (t=6.066), body mass index (t=2.804), disease duration (t=2.552), intraocular pressure (t=2.574), high-density lipoprotein (t=2.729), fasting blood glucose (t=2.022), glycosylated hemoglobin (t=7.235), SDF-1 level (t=14.319), and internal carotid artery stiffness index (t=2.633) were higher than those in the NAION group, while diastolic blood pressure was lower (P＜0.05). ROC curve analysis showed that the AUC of SDF-1 level combined with internal carotid artery stiffness index in predicting the risk of adverse prognosis was 0.894 [95% confidence interval (CI) 0.803-0.945], with a sensitivity of 87.98% and a specificity of 95.69%. Logistic regression analysis demonstrated significant independent correlations between SDF-1 level (OR=1.682, 95%CI 1.156-1.986), internal carotid artery stiffness index (OR=1.826, 95%CI 1.369-2.648), and the risk of ME in NAION patients (P＜0.05). Subgroup analysis revealed that elevated SDF-1 level and internal carotid artery stiffness index were associated with a higher risk of NAION with ME (Pfor trend＜0.05). RCS analysis demonstrated a nonlinear dose-response relationship between the continuous changes in SDF-1 level and internal carotid artery stiffness index and the risk of NAION with ME (P＜0.05). Mediation effect model analysis showed that internal carotid artery stiffness index played a mediating role between SDF-1 level and the risk of NAION with ME. ConclusionsSDF-1 level and internal carotid artery stiffness index are independent risk factors for ME in NAION patients. The combined detection of these two indicators holds significant value in predicting disease progression.
Objective To find a kind of simple and effective method for purifying and label ing stromal vascular fraction cells (SVFs) so as to provide a theoretical basis for cl inical application of SVFs. Methods The subcutaneous adi pose tissue were harvested form volunteers. The adi pose tissue was digested with 0.065%, 0.125%, and 0.185% type I collagenase,respectively. SVFs were harvested after digestion and counted. After trypan blue staining, the rate of viable cells was observed. SVFs was labeled by 1, 1’-dioctadecyl-3, 3, 3’, 3’-2-tetramethy-lindocyanine perchlorate (DiI). The fluorescent label ing and growth was observed under an inverted fluorescence microscope. MTT assay was used to detect cell proliferation. Results The number of SVFs was (138.68 ± 11.64) × 104, (183.80 ± 10.16) × 104, and (293.07 ± 8.31) × 104 in 0.065% group, 0.125% group, and 0.185% group, respectively, showing significant differences among 3 groups (P lt; 0.01). The rates of viable cells were 91% ± 2%, 90% ± 2%, and 81% ± 2% in 0.065% group, 0.125% group, and 0.185% group, respectively, and it was significantly higher in 0.065% group and 0.125% group than in 0.185% group (P lt; 0.01), but no significant difference was found between 0.065% group and 0.125% group (P=0.881). Inverted fluorescence microscope showed that the cell membranes could be labeled by DiI with intact cell membrane, abundant cytoplasm, and good shape, but nucleus could not labeled. SVFs labeled by DiI could be cultured successfully and maintained a normal form. MTT assay showed that similar curves of the cell growth were observed before and after DiI labeled to SVFs. Conclusion The optimal collagenase concentration for purifying SVFs is 0.125%. DiI is a kind of ideal fluorescent dye for SVFs.
Objective To investigate the role and relative mechanism of stromal cell derived factorl (SDF-1) secreted by nucleus pulposus cells (NPCs) on the proliferation of vascular endothelial cells (VECs). Methods The NPCs were isolated from the degenerated disc specimens after discectomy. NPCs at passage 1 were transfected with lentivirus-mediated SDF-1 over-expression; transfected and untransfected NPCs at passage 2 were cultured in the three-dimensional alvetex® scaffold, then they were co-cultured with HMEC-1 cells. The morphology of NPCs was observed by scanning electron microscope (SEM), and the apoptosis of HMEC-1 cells was detected by Annexin V/propidiumiodide staining after 72 hours co-culutre. The proliferation of HMEC-1 cells was detected by cell counting kit 8 at 12, 24, 48, and 72 hours in transfected group and untransfected group, respectively. ELISA was used to measure the vascular endothelial growth factor (VEGF) expression level. The virus transfection efficiency and relative Akt pathway were determined by Western blot. Results The NPCs maintained cell phenotype and secreted much extracellular matrix in three-dimensional-culture by SEM observation. In the co-culutre system, after NPCs were transfected with SDF-1 over-expression lentivirus, the proliferation of HMEC-1 cells was significantly increased, while the apoptosis was decreased obviously. The ELISA results demonstrated that the amount of VEGF was remarkably increased in the culture medium. Furthermore, SDF-1 promoted the up-regulation of phosphorylate Akt expression; after inhibition of Akt expression by GSK690693, the proliferation rate of VECs decreased significantly. Conclusion Over-expression of SDF-1 by NPCs is beneficial for VECs proliferation, which is involved in SDF-1-Akt signalling pathway.
ObjectiveTo explore effect and mechanism of the carcinoma associated fibroblasts (CAFs) of breast cancer on growth and metastasis of breast cancer induced in nude mice by inoculation of CAFs and breast cancer cells. MethodsBreast cancer cell line of MDA-MB-231 (abbreviated as MDA), CAFs, and normal breast tissue fibroblasts (NFs) of the same breast cancer patient were collected, and mixed with normal saline (NS) or SDF-1 ligand blockers of four nitrogen heterocyclic fourteen alka (AMD3100, abbreviated as AMD) for inoculation of nude mice in vivo. According to the different combination, 36 nude mice were randomly divided into 6 groups:MDA+NS group, NFs+NS group, MDA+NFs+NS group, MDA+NFs+AMD group, MDA+CAFs+AMD group, and MDA+CAFs+NS group. Forty six days after the inoculation and feeding, volume of tumor, metastasis of lymph node, lung or liver were observed. In addition, level of plasma SDF-1 was tested by using ELISA method, and expressions of SDF-1 mRNA and protein in tumor specimens were detected by using real-time PCR and Western blot method respectively. ResultsExcept for NFs+ NS group, implanted tumor could be seen in nude mice of other 5 groups. In MDA+CAFs+NS group, the volume of tumor[(9.092±2.662) cm3], level of plasma SDF-1[(75.25±16.23) ng/L], and expression levels of SDF-1 mRNA (the median level was 14.714) and protein (the median level was 0.673). of tumor tissue were significantly greater or higher than those of the other 5 groups (P < 0.050). In addition, lymph node metastasis were found in 4 mice in MDA+CAFs+NS group, and 2 in MDA+NS group. The tumor metastasis of lung and liver was not found in all nude mice. ConclusionsCAFs can promote growth and lymph node metastasis of breast cancer, whose mechanism is related with SDF-1 secreted by CAFs and SDF-1/CXC chemokine receptor 4 (CXCR4), signal pathway.
ObjectiveTo review the research progress of tissue engineered scaffolds and stromal-derived factor 1 (SDF-1) composite graft. MethodsThe recent papers about SDF-1 with different kinds of tissue engineered scaffolds were reviewed and analyzed. The primary mechanism of SDF-1 homing function for stem cells was retrospected. The results of different kinds of tissue engineered scaffolds carrying SDF-1 for repairing the injured tissues and organs were reviewed. ResultsIt is shown that SDF-1 combined with tissue engineered scaffolds will play a role of multipotent stem cells chemotaxis, however, the exact chemotaxis mechanism has not been fully understood. It still needs more researches of SDF-1 effects in vivo. ConclusionAlthough some research progress has been made in regeneration in situ of tissue engineered scaffolds combined with SDF-1, it will need to further study on the mechanism of chemotactic functions of SDF-1 and its influence on proliferation and differentiation of cells.