RESUMEN
In order to understand the students'knowledge mastery,assess teaching effects and improve the quality of teaching,this article analyzed the test papers of Neurology among the five-year program clinical medicine students of Grade 2006 in our College.The results indicated that the mean score of the exam was 87.29 with a standard deviation of 7.90.The distribution was negatively skewed.The degree of difficulty was 0.78 and the discrimination is 0.19.The reliability was 0.59.It demonstrated that the test papers had a good reliability,the design of the test papers was too easy and could not reasonablely discriminate the degree in evaluating the students'mastery of the course.Therefore,we shoud pay attention to appropriate difficulty and ensare quality in preparing test papers to scientifically evaluate teaching quality.
RESUMEN
The simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood were carried out using bioreactors. The co-culture of umbilical cord blood (UCB)-derived hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) was performed within spinner flasks and a rotating wall vessel (RWV) bioreactor using glass-coated styrene copolymer (GCSC) microcarriers. The medium used was composed of serum-free IMDM containing a cocktail of SCF 15 ng·mL(-1), FL 5 ng·mL(-1), TPO 6 ng·mL(-1), IL-3 15 ng·mL(-1), G-CSF 1 ng·mL(-1) and GM-CSF 5 ng·mL(-1). Accessory stromal cells derived from normal allogeneic adipose tissue were encapsulated in alginate-chitosan (AC) beads and used as feeding cells. The quality of the harvested UCB-HSCs and MSCs was assessed by immunophenotype analysis, methylcellulose colony and multi-lineage differentiation assays. After 12 days of culture, the fold-expansion of total cell numbers, colony-forming units (CFU-C), CD34(+)/CD45(+)/CD105(-) (HSCs) cells and CD34(-)/CD45(-)/CD105(+) (MSCs) cells using the RWV bioreactor were (3.7 ± 0.3)- , (5.1 ± 1.2)- , (5.2 ± 0.4)- , and (13.9 ± 1.2)-fold respectively, significantly better than those obtained using spinner flasks. Moreover, UCB-HSCs and UCB-MSCs could be easily separated by gravity sedimentation after the co-culture period as only UCB-MSCs adhered on to the microcarriers. Simultaneously, we found that the fibroblast-like cells growing on the surface of the GCSC microcarriers could be induced and differentiated towards the osteoblastic, chondrocytic and adipocytic lineages. Phenotypically, these cells were very similarly to the MSCs derived from bone marrow positively expressing the MSCs-related markers CD13, CD44, CD73 and CD105, while negatively expressing the HSCs-related markers CD34, CD45 and HLA-DR. It was thus demonstrated that the simultaneous expansion and harvest of UCB-HSCs and UCB-MSCs is possible to be accomplished using a feasible bioreactor culture system such as the RWV bioreactor with the support of GCSC microcarriers.
Asunto(s)
Sangre Fetal/citología , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/fisiología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Antígenos CD/metabolismo , Antígenos CD34/metabolismo , Reactores Biológicos , Recuento de Células , Técnicas de Cultivo de Célula , Proliferación Celular , Separación Celular/métodos , Células Cultivadas , Técnicas de Cocultivo/métodos , Endoglina , Células Madre Hematopoyéticas/metabolismo , Humanos , Antígenos Comunes de Leucocito/metabolismo , Células Madre Mesenquimatosas/metabolismo , Receptores de Superficie Celular/metabolismo , Factores de TiempoRESUMEN
Rotating wall vessel (RWV) was used for the ex-vivo expansion of umbilical cord blood stem cells to meet the requirement of clinical application in the aspect of quantity and quality of the stem cells. The mononuclear cells (MNCs) from umbilical cord blood were cultured in T-flasks for 24 h, and then inoculated in RWV to culture for 200 h. The nucleated-cell numbers, pH and osmolality of the culture medium were determined every 24 h. The CD34+ cells content was measured and CFU-GM culture was carried out at 144 h and 197 h. Nucleated cells (NC) and CD34+ cells had a 435.5 +/- 87.6 fold expansion and a 32.7 +/- 15.6 fold expansion respectively in 197 h, and CFU-GM (colony-forming unit-granulocyte/macrophage) cells had a 21.7 +/- 4.9 fold expansion. In the whole course of culture, the pH and osmolality of the medium in the RWV were kept in the optimal hematopoietic stem cells' expansion conditions. pH was kept from 7.2 to 7.4, and the osmolality was kept from 290 mmol/kg to 310 mmol/kg. Owing to its structural particularity, the RWV could ensure cells to grow in the suspension state, could simulate the micro-environment of umbilical cord blood, and thus could make the hematopoietic stem cells expand largely in the RWV in short time.