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 evaluate the mechanism of stromal vascular fraction (SVF) promoting angiogenesis and tissue regeneration in tissue engineering chamber. MethodsTwenty-four 6-month-old New Zealand white rabbits, male or female, weighing 2.5-2.8 kg, were selected. Thoracic dorsal arteriovenous bundle combined with collagen type I scaffold was transplanted to dorsal side, and wrapped by cylindrical hollow silicone chamber; all animals were randomly divided into the experimental group (n=12) and the control group (n=12). SVF was isolated from the back fat pads of rabbits in experimental group and labelled with DiI at 2 weeks after operation. The 1 mL cell suspension (1×106 cells/mL) and equal saline were injected into the chamber in experimental group and control group, respectively. The regenerative tissues were harvested for general observation and HE staining at 2 and 4 weeks after injection;and immunofluorescent staining was carried out in experimental group at 4 weeks. ResultsAt 2 weeks after injection, the regenerative tissue was cylindrical; obvious vessel network and incompletely degradable collagen scaffold could be seen on the surface of the new tissue in 2 groups. The volume of new tissue was (0.87±0.11) mL in experimental group, and (0.72±0.08) mL in control group at 2 weeks, showing significant difference (t=2.701, P=0.011). At 4 weeks, little collagen scaffold could be seen on the surface in control group, but no collagen scaffold in experimental group; the volume of new tissue was (0.74±0.14) mL in experimental group, and (0.64±0.10) mL in control group, showing no significant difference (t=1.424, P=0.093). HE staining showed new mature vessels at 4 weeks, but no adipose tissue or fat lobulus formed in both groups; the capillary density was significantly higher in experimental group than in control group at 2 weeks (t=6.291, P=0.000) and at 4 weeks (t=5.445, P=0.000). The immunofluorescent staining found that SVF survived and located at the edge area after 4 weeks; the expressions of CD31 and DiI were positive in some endothelial cells. ConclusionSVF can promote the angiogenesis and tissue regeneration in tissue engineering chamber, but it can not differentiate into adipocyte spontaneously without adipogenic microenvironment.
The osteogenc potential of bone marrow has been proved by experiment. To investigate more in details, bone marrow was obtained from the trochanteric region of femur of NewZealand rabbit in 4 to 8 weeks old. After being cultured in vitro for one week, the hematopoietic component of the bone marrow had disappeared, thus the stromal cells were obtained. Then the stromal cells were subcultured in cultural fluid containing dexamethasone (10-8 mol/L) and natrium glycerophosphate (10mmol/L). Under the phasecontrast microscope, it was found that being cultured for 15 days. The stromal cells were lined up in one layer and late the secretion activity was increased and gradually transformed into multilayer structure and was congregated into diffused opaque clusters in twenty days. During culture, the cells were examined by tetracycline fluorescence label, histochemistry stains, transmission electron microscopy, scanning electron microscopy and energy dispersive X-ray microanalysis. The results showed that the morphological and biological characteristics of the cultured stromal cells derived from the bone marrow were similiar to those of osteoblasts and could synthesized mineralized new bone tissue in vitro.
ObjectiveTo explored the effect of stromal cell-derived factor 1α (SDF-1α) on promoting the migration ability of rat adipose derived stem cells (rADSCs) by constructed the rADSCs overexpression SDF-1α via adenovirus transfection.MethodsrADSCs were isolated from adipose tissue of 6-week-old SPF Sprague Dawley rats. Morphological observation, multi-directional differentiations (osteogenic, adipogenic, and chondrogenic inductions), and flow cytometry identification were performed. Transwell cell migration experiment was used to observe and screen the optimal concentration of exogenous SDF-1α to optimize the migration ability of rADSCs; the optimal multiplicity of infection (MOI) of rADSCs was screened by observing the cell status and fluorescence expression after transfection. Then the third generation of rADSCs were divided into 4 groups: group A was pure rADSCs; group B was rADSCs co-cultured with SDF-1α at the best concentration; group C was rADSCs infected with recombinant adenovirus-mediated green fluorescent protein (Adv-GFP) with the best MOI; group D was rADSCs infected with Adv-GFP-SDF-1α overexpression adenovirus with the best MOI. Cell counting kit 8 (CCK-8) and Transwell cell migration experiment were preformed to detect and compare the effect of exogenous SDF-1α and SDF-1α overexpression on the proliferation and migration ability of rADSCs.ResultsThe cell morphology, multi-directional differentiations, and flow cytometry identification showed that the cultured cells were rADSCs. After screening, the optimal stimulating concentration of exogenous SDF-1α was 12.5 nmol/L; the optimal MOI of Adv-GFP adenovirus was 200; the optimal MOI of Adv-GFP-SDF-1α overexpression adenovirus was 400. CCK-8 method and Transwell cell migration experiment showed that compared with groups A and C, groups B and D could significantly improve the proliferation and migration of rADSCs (P<0.05); the effect of group D on enhancing the migration of rADSCs was weaker than that of group B, but the effect of promoting the proliferation of rADSCs was stronger than that of group D (P<0.05).ConclusionSDF-1α overexpression modification on rADSCs can significantly promote the proliferation and migration ability, which may be a potential method to optimize the application of ADSCs in tissue regeneration and wound repair.
Objective To introduce the related issues in the clinical translational application of adipose-derived stem cells (ASCs). Methods The latest papers were extensively reviewed, concerning the issues of ASCs production, management, transportation, use, and safety during clinical application. Results ASCs, as a new member of adult stem cells family, bring to wide application prospect in the field of regenerative medicine. Over 40 clinical trials using ASCs conducted in 15 countries have been registered on the website (http://www.clinicaltrials.gov) of the National Institutes of Health (NIH), suggesting that ASCs represents a promising approach to future cell-based therapies. In the clinical translational application, the related issues included the quality control standard that management and production should follow, the prevention measures of pathogenic microorganism pollution, the requirements of enzymes and related reagent in separation process, possible effect of donor site, age, and sex in sampling, low temperature storage, product transportation, and safety. Conclusion ASCs have the advantage of clinical translational application, much attention should be paid to these issues in clinical application to accelerate the clinical translation process.
Objective To investigate the influence on matrix metalloproteinases (MMP) 3, 9, and 13 levels of human articular cartilage cells after blocking stromal cell derived factor 1 (SDF-1)/ chemokine receptor 4 (CXCR4) signaling pathway withAMD3100 and to define the function mechanism of AMD3100. Methods A total of 144 cartilage blocks from 12 osteoarthritis (OA) patients undergoing total knee arthroplasty (OA cartilage group) and 144 normal cartilage blocks (Mankin score of 0 or 1) from 12 patients undergoing traumatic amputation (normal cartilage group). OA cartilage group was further divided into subgroups A1, B1, and C1, and normal cartilage group into subgroups A2, B2, and C2. The cartilage tissues were cultured in DMEM solution containing 100 ng/mL SDF-1 and 1 000 nmol/L AMD3100 in subgroup A, 100 ng/mL SDF-1 and 1 000 nmol/L MAB310 in subgroup B, and 100 ng/mL SDF-1 in subgroup C, respectively. The levels of MMP-3, 9, and 13 were measured by ELISA; the expressions of MMP-3, 9, and 13mRNA were tested by RT-PCR. Results ELISA and RT-PCR results showed that the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly lower in subgroup A than in subgroups B and C at the same time points (P lt; 0.05); the levels of MMP-3, 9, and 13 and the expressions of MMP-3, 9, and 13 mRNA were significantly higher in OA cartilage group than in normal cartilage group at the same time points (P lt; 0.05). Conclusion SDF-1 could induce overexpression and release of MMP-3, 9, and 13 in the articular cartilage through the SDF-1/CXCR4 signaling pathway; AMD3100 could reduce the mRNA expressions and secretion of MMP-3, 9, and 13 in OA cartilage by blocking the SDF-1/CXCR4 signaling pathway; but AMD3100 could not make the secretion of MMP-3, 9, and 13 return to normal levels in OA cartilage.
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.