Bone marrow-derived stromal cells and cartilage tissue engineering     

CHEN Wen-fang  WANG Lian-tang 《园艺学报》2003,19(2):279-283

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.  相似文献   

Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines     

CHEN Dong-ying  ZHU Quan-sheng  LIU Chao  QIU Ju-shi 《园艺学报》2003,19(6):751-756

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.  相似文献   

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro     

CAI Yun-feng  MIN Jun  FANG Tian-ling  CHU Zhong-hua  DENG Xiao-geng  HE Jing-song  CHEN Ji-sheng 《园艺学报》2003,19(8):1029-1033

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.  相似文献   

Biological characteristics of long passaging rat mesenchymal stem cells     

LEI Jun-xia  LI Hao-wei  HUANG Chun-nong  ZHU Mei-ling  WEN Guan-mei  ZHANG Xiu-ming  LI Yan  LI Shu-nong 《园艺学报》2003,19(10):1325-1330

AIM:To investigate multi-potential of rat bone marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC were long passaged in vitro. METHODS:Cellular cycles of different passages were assayed by FACSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS:The early passages and long-term passages all showed strong proliferation; passage 6, passage 25 and passage 45 all showed normal karyotype. CONCLUSION:Long-term culture and passage of rBMMSC still remains strong proliferation. With this capability, the mutation inclination is not enhanced.  相似文献   

Adult rat and human bone marrow mesenchymal stem cells differentiate into neurons with Musk's polypeptide     

XIAO Qing-zhong  LI Hao-wei  WEN Guan-mei  HUANG Shao-hua ZHANG Xiu-ming  LI Yan  LI Shu-nong 《园艺学报》2002,18(10):1179-1182

AIM: To investigate the differentiation from adult rat and human bone marrow mesenchymal stem cells (BMMSCs) into neuron with musk polypeptide (Mu-P).METHODS: Adult rat and human BMMSCs were induced with Mu-P.Neuron-specific enolase (NSE),neurofilament (NF),Nestin,glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry.RESULTS: Simple methods with Mu-P induced adult rat and human BMMSCs exhibiting a neuronal phenotype,expressing Nestin at 3 hours to 5 hours,and expressing NE and NF at 5 hours to 7 days.But the neuron-like cells didn't express the glial astrocyte marker GFAP.CONCLUSION: Adult rat and human BMMSCs can be induced to differentiate into neurons with Mu-P.  相似文献   

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts     

ZHANG Li-rong  XIANG Peng  XIA Wen-jie  CHEN Zhen-guang  ZHANG Xiu-ming  LI Yan  LI Shu-nong 《园艺学报》2002,18(8):896-899

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.  相似文献   

Bone marrow mesenchymal stem cells are induced to differentiate into cardiomyocytes by hepatocyte growth factor in vitro     

FANG Tian-ling  MIN Jun  DENG Xiao-geng  QIAN Shi-kun  CHU Zhong-hua  CHEN Ya-jin  SHAO Jing  WEI Jing  CHEN Ji-sheng 《园艺学报》2004,20(7):1167-1170

AIM: To explore a new method of hepatocyte growth factor (HGF) inducing bone marrow mesenchymal stem cells (MSC) to differentiate into cardiomyocytes. METHODS: Bone marrow MSC was cultured with DMEM media (10% fetal calf serum) 4-6 passages, and induced by HGF (10 μg/L) for 30 d. Automatical beating of the differentiated cells was observed daily with transverse microscopy, or under condition of 0.1% isoproterenol or cal-cium-deprived incubation. Specific cardiac myosin in the cells was indentified by immunochemistry. RESULTS: At 14-20 d of differentiation, bone marrow mesenchymal stem cells formed clones, in 10%-50% of which spontaneous beating cell-mass had come to continuously exist. Isoproterenol increased the beating rate and calcium-deprived media inhibited the beating. The cells were identified to be cardiomyocytes by expression of cardiac myosin heavy chain. CONCLUSION: HGF may induce bone marrow mesenchymal stem cells into cardiomyocytes with high efficiency, but the differentiating pathway of stem cells remains to be further studied.  相似文献   

Are dual-purpose and male layer chickens more resilient against a low-protein-low-soybean diet than slow-growing broilers?     

M. Kreuzer  S. Müller  L. Mazzolini  R. E. Messikommer  I. D. M. Gangnat 《British poultry science》2020,61(1):33-42

ABSTRACT

1. Although fattening dual-purpose types or male layer hybrid chickens appears more ethical than the common practice of culling day-old male layer chicks, the lower feed efficiency of these birds raises concerns. Replacing feed ingredients that compete with food production by those of lower value for human nutrition would be beneficial.

2. Lohmann Dual (LD), a modern dual-purpose type, Lohmann Brown (LB), a male layer hybrid, and Hubbard JA 957 (HU), a slow-growing broiler type, were fattened for nine weeks on two diets (control or ?20% crude protein; n = 6 × 12 birds). Growth, carcass and meat quality were analysed.

3. Growth performance of HU exceeded that of LD and especially of LB. The growth depression caused by the low-protein diet fed to LD (?7%) was only half of that found in HU (?13%). The LD fed the control diet had the same feed efficiency as the HU fed the low-protein diet. Even the LB had a lower performance and feed efficiency with the low-protein diet in growth. There was a gradient in carcass properties (weight, dressing percentage, breast meat yield, breast proportion and breast angle) from HU to LD to LB, with some additional adverse effects of the low-protein diet especially in HU. There were some breed differences in fatty acid profile in the intramuscular fat.

4. In conclusion, the dual-purpose type used complied with regulations for Swiss organic poultry systems in terms of growth rate and was found to respond less when fed a low-protein diet than the slow-growing broiler type. The LB males were inferior in all growth and carcass quality traits. Future studies need to determine the exact protein and amino acid requirements of dual-purpose and layer hybrid chickens and the economic feasibility of the systems, especially for organic farming.  相似文献   

工业氟污染对山羊硬组织损害的研究   总被引：5，自引：1，他引：5  

王俊东  陈友发 《畜牧兽医学报》1992,23(2):141-145

40只6～10月龄本地山羊在氟重污染牧场放牧18个月,对逐月牧草水溶氟,不同时期骨氟、枯草期血钙及碱性磷酸酶作了测定。同时应用X射线及扫描电镜对下颌、肋骨与牙齿结构作了观察。结果指出,随着骨氟增加,血钙降低、碱性磷酸酶升高、骨皮质变薄、密度降低、骨表面疏松多孔,牙齿磨灭不齐、钙化不良、釉质与本质结构改变。干早气候是枯草期牧草氟升高的先决条件,枯草期牧草高氟与钙的负平衡是形成骨质疏松的主要原因,而青草或与枯草期牧草氟含量及动物缺钙程度的不同是引起牙齿磨灭不齐、缩短寿命的关键因素。  相似文献   

Erythroid progenitor cells from pig bone marrow and peripheral blood     

Kovacević M  Bozić T  Ivanović Z 《Veterinary journal (London, England : 1997)》1999,158(3):143-203

With the intention of using the pig as a large animal model in haematopoietic research, a clonal assay in methylcellulose was developed and the optimal conditions for raising erythroid progenitors from adult pig bone marrow (BM) and peripheral blood (PB) have been established. Progenitor cells were stimulated to proliferate and differentiate in vitro by growth factors containing leucocyte condition medium (LCM), and with recombinant human erythropoietin (rhEpo). The number of PB BFU-E (burst forming units - erythroid) directly depended on the concentration of LCM, but BM BFU-E were not dependent on LCM. Both CFU-E (colony forming units - erythroid) and BFU-E were rhEpo dependent. Despite relatively high but expected individual variations, the mean number of colonies, as well as the functional characteristics of progenitor cells investigated, were similar to those of miniature pigs and some other mammals.  相似文献