Objective To study the biological behavior of osteoblast and vascular endothelial cell culture. Methods The osteoblasts and vascular endothelial cells were obtained from calvarial bone and renal cortox of 2-week rabbits respectively. The experiment were divided into group A (osteoblasts), group B (vascular endothelial cells) and group C(co-cultured osteoblasts and vascular endothelial cells). The cells were identified with cytoimmunochemical staining. The cellular biological behavior and compatibilitywere observed under inverted phase contrast microscope and with histological staining. The cells viability and alkaline phosphatase(ALP) activity were measured. Results The cytoimmunochemical staining showed that the cultured cells were osteoblasts and vascular endothelial cells .The cellular compatibility of osteoblasts and vascular endothelial cells was good. The ALP activity was higher in group C than in group A and group B(P＜0.01), and it was higher in group A than in group B(P＜0.05). In group C, the cellproliferation were increased slowly early, but fast later. Conclusion Thecellular compatibility of osteoblasts and vascular endothelial cells were good. The vascular endothelial cells can significantly increased the osteoblast viability and ALP activity,and the combined cultured cells have greater proliferation ability.
OBJECTIVE This paper was to study the biological characteristics of the transformed human embryonic tendon cells, the relation between cell growth and culture conditions, and to compare these features with that of human embryonic tendon cells. METHODS The pts A58H plasmid had successfully used to transform a tendon cell line from human embryo in our past work. The human embryonic tendon cells and the transformed human embryonic tendon cells were cultured in vitro. In different culture conditions, the growth curve were drawn respectively. Population dependence and proliferation capability of the cells were investigated through plate cloning test and soft agar culture. The collagen secreted by cells was identified by immunohistochemical method. RESULTS In routine culture condition, the growth properties of the human embryonic tendon cell and transformed cells were almost identical. The growth properties of the transformed cells were not changed when the cells were frozen storage. There were changes of growth characteristics of the transformed cells when the culture temperature was changed. The transformed cells could subcultured continually and permanently. The proliferation capability of the transformed cells were ber than that of the human embryonic tendon cells. Moreover, the growth of the transformed cells was serum-dependent, and the phenomenon of contact inhibition was observed. The transformed cells were not able to grow on soft agar culture. They had the capacity of secreting collagen type I. CONCLUSION The transformed human embryonic tendon cells could be subcultured continually and permanently, and their growth could be controlled by changing their culture conditions and they had no malignant tendency in biological characteristics. They could be taken as an ideal experimental material for tendon engineering.
摘要:目的: 金黄色葡萄球菌(金葡菌)的感染近年来已成为医院内的主要致病菌,而其耐药性也呈逐渐升高的趋势,为了解该菌在我院的感染和耐药情况,为临床合理使用抗生素提供科学依据。 方法 : 用经典生理生化鉴定方法,对各种临床标本主要来源于痰液和各种伤口脓液标本分离到的102株金葡菌进行生物学特性及药敏试验。 结果 : 从我们医院2007年5月至2009年8月所分离出来的102株金葡菌中青霉素耐药性8923%,氨苄青霉素耐药率为9385%,没有发现万古霉素耐药菌。 结论 : 除万古霉素外,耐药率较低的依次是利福平、苯唑青霉素、环丙沙星、呋喃妥因、阿米卡星、磺胺甲基异恶唑、红霉素,而青霉素G、氨苄青霉素、四环素耐药性情况非常严重,并且多重耐药,耐药性强,应引起临床的高度重视。Abstract: Objective: To analyze the bionomics and antimicrobial susceptibility of staphylococcus aureus, which was the main pathogenic bacterium with high drug tolerance in our hospital, in order to provide the rational use of antibiotics. Methods : Samples of one hundred and two staphylococcus aureus cases from sputamentum and pus were evaluated by classic physiology and biochemistry methods to test the bionomics and antimicrobial susceptibility. Results : The drug resistance rate to penicillin, penbritin and vancomycin was 8923%, 9385% and 0, separately. Conclusion : Besides vancomycin, the drug resistance rate of rifampicin, oxazocilline, ciprofloxacin, furadantin, amikacin, sulfamethoxazole and sulfamethoxazole increased one by one. The resistance to penicillin G, penbritin and tetracycline was serious, including multidrug resistant, which should be paid highly attention.
Objective To explore a method to isolate, culture and multiplicate the placentaderived mesenchymal stem cells (PMSCs) and the bone marrow-derived mesenchymal stem cells (BMSCs) of rabbit，and to compare their biological characteristics. Methods PMSCs were isolated from placenta of 1fetation rabbitby Percoll density gradient centrifuge and cultured in vitro. BMSCs were isolated from hindlimb bone marrow blood of 1 new born rabbit by direct plates culturemethod. The 3rd passage PMSCs and BMSCs were observed by inverted phase contrast microscope. The stem cell marker (CD44, CD105, CD34 and CD40L) were examined by immunohistochemistry. The 2nd passage PMSCs and BMSCs were co-cultured with biomaterials，(1.0-1.5)×106 cells in one biomaterial， and then observed by aematoxylinstaining after 5 days,and by SEM after 3 days and 8 days. Results PMSCs and BMSCs were both uniformly spondle-shaped in appearance and showed active proliferative capacity. The proliferative ability of PMSCs were quite b and declined with passages. After cultured 10 passages in vitro, its growthslowed. Both PMSCs and BMSCs expressed CD44 and CD105,but did not express CD34 and CD40L immunoreactivity. PMSCs and BMSCs poliferated and adhered to the surface of biomaterials, and cell formed clumps and network; the cells proliferation and the matrix were seen in the pore after 5 days of culture. The observation ofSEM showed that many cells adhered to the biomaterials with spindle-shape and polygon after 3 days; and that PMSCs and BMSCs grew,arranged in layers andsecreted many matrices; the reticular collagen formed arround cells after 8 days. Conclusion PMSCs and BMSCs have similar biological characteristics and PMSCs can be served as excellent seedingcells for tissue engineering.
Following the peritendon was removed by means of microsurgical technique, the tenocyte was isolated from the human embryonic tendons by digesting it with trypsin and collagenase. These cells were all stored in frozen condition until they were cultured by F12 culture fluid added with 20% FBS to the 15th generation.These cells were able to grow adhering to the wall and stop growing with contact inhibition. The time of cellsgroup duplication was 4 days, which was similar to the peak time of its mitosis. The number of its chromosome group 2n=46 was 87.5-91.0%. The optimal conditions for tendon cell culture in vitro were investigated, and it was found that after they were reaminated and subcultured the frozen storage didn’t influence their growth, morphology, genetic characteristics. In our research we detected the content generation cells and found the cultured human embryonic tenocyte had same ability never changed with the cells subcultured. We also disscussed the future of tenocyte-a biomaterial in the field of artificial implant.
Objective To summarize the research progress of biological characteristics and advantages of Wharton’s jelly-mesenchymal stem cells (WJ-MSCs). Methods The related l iterature on the biological characteristics of WJ-MSCs,umbil ical cord blood MSCs (UBMSCs) and bone marrow MSCs (BMSCs) was extensively reviewed and analyzed. Results A large number of MSCs which are able to self-repl icate, self-renew and have high prol iferation and multipotent differentiation can be isolated from the Wharton’s jelly of umbil ical cord. WJ-MSCs have many advantages in isolation time, isolation efficience, expansion time, passage capacity, expansion capacity when compared with UBMSCs and BMSCs. Conclusion WJ- MSCs have numerous advantages of convenient and abundant sources, relatively pure, non-ethical issues, and so on, which can be used for cell transplant therapy, gene therapy, and the ideal seed cells of building tissue engineered organ, so they provide new ideas for tissue regeneration repair and reconstruction.
ObjectiveTo summarize the research status and biological characteristics of stromal fibroblast in breast cancer. MethodsRelevant literatures about the breast cancer stromal fibroblasts published recently were collected and reviewed. ResultsIn addition to cancer cells, breast cancer included stromal cells. The fibroblasts were the major components of breast cancer stromal, which had significantly different biological characteristics from normal fibroblasts. The fibroblasts were characterized by α-SMA positive, p53 gene mutation, secretion of various cytokines or chemokines in addition to the production of collagen substances, involving in breast cancer growth, migration, invasion and metastasis through a variety of signaling pathways. ConclusionThe biological characteristics of stromal fibroblasts in breast cancer may reflect lesion properties, be of great importance to diagnosis and differential diagnosis and prognosis prediction of breast cancer. More attentions will be paid to the target therapy for stromal fibroblasts in breast cancer.
Objective To investigate the biological and biomechanical characteristics of acellular porcine aortic valve with dye mediated photo oxidation so that a new and better bioprosthetic valve materials can be obtained. Methods Thirty porcine aortic valves were divided into three groups with random number table. Acellular valves (n=10) were stabilized by dye mediated photo oxidation in dye mediated photo oxidation group; acellular valves (n=10) were stabilized by glutaraldehyde in glutaraldehyde group; and acellular valves (n=10) were acellularized only in acellular valves group. Thickness, appearance, histology, water content, shrinkage temperature, breaking strength and soluble protein level of acellular porcine aortic in three groups were tested respectively. Results There were light blue, soft, flexible and unshrinking valves in dye mediated photo oxidation group. Compared to valves in glutaraldehyde group, valves in dye mediated photo oxidation group had lighter thickness（0.26±0.09mm vs. 0.38±0.08mm，Plt;0.05）, more water content（86.30%±4.03% vs. 71.10%±3.23%，Plt;0.05）, and lower shrinkage temperature （76.30±0.70℃ vs. 87.70±0.30℃，Plt;0.05）; while these indexes had no statistically significant differences compared to those in acellular valves group. At the same time, compared to valves in acellular valves group, valves in dye mediated photo oxidation group had more breaking strength（17.33±2.65 mPa vs. 9.11±0.95 mPa，Plt;0.05） and lower soluble protein level（0.039%±0.013% vs. 0.107%±0.024%，Plt;0.05）; while these indexes had no statistically significant differences compared to those in glutaraldehyde group. Conclusion Acellular porcine aortic valve stabilized by dye mediated photo oxidation has nice biological and biomechanical characteristics.
Objective To observe the effects of cryopreservation and resuscitation on the biological characteristics of mesenchymal stem cells (MSCs) derived f rom human umbilical cord blood. Methods MSCs were isolated and cultured f rom human umbilical cord blood in vitro. The cells were passaged , and the third generation of MSCs were cryopreserved in－196 ℃ liquid nitrogen for 4 weeks with cryopreservation medium , which contained 10 % dimethyl sulfoxide (DMSO) and 90 % fetal calf serum ( FCS) . The morphology , proliferation and differentiation of MSCs were investigated and compared with those of MSCs before cryopreservation. Results There was no significant difference of morphology between pre-cryopreserved MSCs and the ones af ter resuscitation. It was observed that all MSCs were spindle-shaped and showed adherence growth characteristic before and af ter cryopreservation. The cell growth curves of MSCs were also similar before and af ter cryopreservation. Even though the curve of resuscitated MSCs descended a little as compared with that of pre-cryopreserved MSCs , there was no significant difference ( Pgt; 0. 05) . After 2-week adipocytic differentiation induction , fat drops could be found in the kytoplasm of MSCs and they were red when stained with oil-red O staining , which suggested that MSCs could be induced and differentiated into adipocytes. Af ter 4-week osteoblastic differentiation induction , MSCs could be induced and differentiated into osteoblasts , and calcium node showed black when stained with Von Kossa staining. There were no significant changes of the differentiating ability of MSCs into adipocyte and osteoblast before and after cryopreservation. Conclusion MSCs derived from human umbilical cord blood maintains their biological characteristics af ter cryopreservation and resuscitation.