Effect of siwutang on recovery of anemia induced by chemotherapy in mice and its mechanism / 吉林大学学报(医学版)

Objective: To study the recovery effect of Siwutang on the anemia induced by chemotherapy in the mice, and to clarify its mechanism. Methods: A total of 24 mice were randomly divided into control group (n=8, not given any drugs), model group (n= 8, given 5-fluorouracil), Siwutang group (n= 8, given 5-fluorouracil + Siwutang). After administration for 15 d, the peripheral blood of mice was taken, and the morphology and the number of red cells were observed under microscope. The number of peripheral blood red cells and the levels of hemoglobin of the mice in various groups were calculated by full-automatic hemocytometer; Double staining was used to mark the surface antigens of red cells, then antibody binding and staining were performed. Flow cytometry was used to detect the number of peripheral blood-labelled and spleen-labelled red cells. Results: Compared with control group, the number of red cells in peripheral blood of the mice in model group was significantly decreased (P<0. 05), and the cells shrinkaged. Compared with model group, the number of red cells in peripheral blood of the mice in Siwutang group was significantly increased (P<0. 05) and the cells showed rounding. Compared with control group, the level of hemoglobin in peripheral blood of the mice in model group was significantly decreased (P<0. 05); compared with model group, the level of hemoglobin in peripheral blood of the mice in Siwutang group was significantly increased (P<0. 01). The flow cytometry results showed that compared with control group, the expression level of CD71/Terll9 in peripheral blood of the mice in model group was significantly decreased (P<0. 01), and the number of red cells in various quadrants was decreased (P<0. 01); compared with model group, the expression level of CD71/Terll9 in peripheral blood of the mice in Siwutang group was significantly increased (P<0. 01), and the number of red cells in various quadrants was increased (P<0. 01); compared with control group, the expression level of CD71/Terll9 in spleen blood of the mice in model group was significantly decreased (P< 0. 01), and the number of cells in various quadrants was decreased (P<0. 01); compared with model group, the expression level of CD71/Terll9 in spleen blood of the mice in Siwutang group was significantly increased (P<0.01), the number of red cells in UL quadrant was decreased (P<0. 01), and the number of red cells in UR and LR quadrants was increased (P<0.01). Conclusion: Siwutang can promote the recovery of anemia induced by chemotherapy in the mice, increase the number of red cells, improve the morphology of red blood cells, increase the level of hemoglobin in peripheral blood, and increase the expression levels of marker antigens on the surface of red cells in the peripheral blood and spleen.

Células madre: buscando marcadores de superficie celular que predispongan compromiso de diferenciación cardiaca / Stem cells: searching predisposition to cardiac commitment by surface markers expression

Resumen Actualmente las enfermedades cardiovasculares se han convertido en un serio problema para los sistemas de salud de todo el mundo, ya que son la principal causa de muerte y representan una enorme carga económica. Este problema ha sido abordado con diferentes estrategias, entre ellas con la ayuda de terapia celular, aunque sin resultados contundentes. Durante más de 20 años, se ha utilizado una gran variedad de células madre en diferentes modelos de infarto del miocardio. El uso de células madre cardiacas (CSC) parece ser la mejor opción, pero la inaccesibilidad y la escasez de estas células hacen que su uso sea muy limitado. Además, existe un riesgo elevado pues tienen que obtenerse directamente del corazón del paciente. A diferencia de las CSC, las células madre adultas derivadas de médula ósea o tejido adiposo, entre otras, representan una opción atractiva debido a su fácil accesibilidad y abundancia, pero sobre todo a la probable existencia de progenitores cardiacos entre sus diferentes subpoblaciones. En esta revisión hacemos un análisis de los marcadores de superficie presentes en CSC en comparación con otras células madre adultas, y sugerimos la preexistencia de células que comparten marcadores de superficie específicos con CSC, la presencia de un inmunofenotipo predecible, aunque en proporciones bajas, pero con un potencial de diferenciación cardiaca similar a las CSC, lo cual podría aumentar su valor terapéutico. Este estudio revela las nuevas perspectivas con respecto a la presencia de dichos marcadores, los cuales comprometerían algunas de estas subpoblaciones a diferenciarse a tejido cardiaco.

Abstract It is well-known that cardiovascular diseases are the leading cause of death world- wide, and represent an important economic burden to health systems. In an attempt to solve this problem, stem cell therapy has emerged as a therapeutic option. Within the last 20 years, a great variety of stem cells have been used in different myocardial infarction models. Up until now, the use of cardiac stem cells (CSCs) has seemed to be the best option, but the inaccessibility and scarcity of these cells make their use unreliable. Additionally, there is a high risk as they have to be obtained directly from the heart of the patient. Unlike CSCs, adult stem cells originating from bone marrow or adipose tissue, among others, appear to be an attractive option due to their easier accessibility and abundance, but particularly due to the probable existence of cardiac progenitors among their different sub-populations. In this review an analysis is made of the surface markers present in CSCs compared with other adult stem cells. This suggested the pre-existence of cells sharing specific surface markers with CSCs, a predictable immunophenotype present in some cells, although in low proportions, and with a potential of cardiac differentiation that could be similar to CSCs, thus increasing their therapeutic value. This study highlights new perspectives regarding MSCs that would enable some of these sub-populations to be differentiated at cardiac tissue level.

In vivo Evaluation of Human Embryonic Stem Cells Isolated by 57-C11 Monoclonal Antibody

BACKGROUND: The normal cells derived from human embryonic stem cells (hESCs) are regarded as substitutes for damaged or dysfunctional adult cells. However, tumorigenicity of hESCs remains a major challenge in clinical application of hESC-derived cell transplantation. Previously, we generated monoclonal antibody (MAb) 57-C11 specific to the surface molecule on undifferentiated hESCs. The aim of this study is to prove whether 57-C11-positive hESCs are pluripotent and tumorigenic in immunodeficient mice. METHODS: Undifferentiated hESCs were mixed with retinoic acid (RA)-differentiated hESCs at different ratios prior to 57-C11-mediated separation. To isolate 57-C11-positive hESCs from the mixture, biotinylated 57-C11 and streptavidin-coated magnetic beads were added to the mixture. Unbound 57-C11-negative hESCs were first isolated after applying magnet to the cell mixture, and 57-C11-bound hESCs were then released from the magnetic beads. In order to measure the efficiency of separation, 57-C11-positive or -negative hESCs were counted after isolation. To evaluate the efficiency of teratoma formation in vivo, 57-C11-positive or negative cells were further injected into left and right, respectively, testes of nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. RESULTS: Approximately 77~100% of undifferentiated hESCs were isolated after applying 57-C11-coated magnetic beads to the mixed cell populations. Importantly, teratomas were not observed in NOD/SCID mice after the injection of isolated 57-C11-negative hESCs, whereas teratomas were observed with 57-C11-positive hESCs. CONCLUSION: 57-C11-positive hESCs are pluripotent and tumorigenic. The combination of 57-C11 and magnetic beads will be useful to eliminate remaining undifferentiated hESCs for the safe cell transplantation.

Effect of different concentrations of fetal bovine serum on bone marrow mesenchymal stem cell purity and cycle / 安徽医科大学学报

Objective To compare the influence of three kinds of complete media with 0. 10 ,0. 15 ,0. 20 fetal bo-vine serum( FBS) on purity and cycle of rat bone marrow mesenchymal stem cells ( BMSCs) cultured in vitro and to seek suitable FBS concentration for the cultivation of the stem cells. Methods SD rats were executed by cervical dislocation method and used whole bone marrow adherence to isolate rat BMSCs. Experiment was divided into A, B,C,3 groups. Compared the expression of CD45, CD29, CD90, CD44 in the three groups in 2,3,4,5 passages ( P2 , P3 , P4 , P5 );the cells of P3 in group A were digested and cultured in three different concentration of com-plete culture media for four days, measuring cell cycle in 24,48,72,96 h by flow cytometry instrument. BMSCs of P3 were collected and inoculated to 6 pieces of 96-well plates, then vaccinated with complete media with 0. 10, 0. 15,0. 20 FBS in every plate, one culture plate was taken out for optical density(OD) meaturement every day with CCK-8. Results CD45 was negative, CD29, CD90, CD44 were positive. The difference of BMSCs surface markers cultured in the three kinds of complete media was bigger in the first two passages, but the difference was less in P3 , P4 , all could obtain pure BMSCs in P4 relatively;according to the results of the cell cycle at the same time, G0/G1 phase:with the increase of concentration of fetal bovine, G0/G1 phase reduced and had no diference in A, B, C groups;G2/M phase:there was difference between them after 24 h(P<0. 05,F=12. 412), but with the extension of time, the differences disappeared;S phase:there was no difference in the three groups;S+G2/M phase increased with the concentration of FBS. According to the result of cell vitality, the OD of ABC three groups increased in turn in 24 h, and there was difference(P<0. 05,F=5. 002), but with the extension of time, there was no obvious difference between them. Conclusion The three kinds of culture media in P4 can obtain pure BM-SCs, cell cycle and vitality show three complete media can promote the growth of BMSCs. There is no difference between them. Culture media with 0. 10 FBS can satisfy the isolation and amplification of BMSCs, in order to ob-tain pure BMSCs in the short term, using culture media with 0. 15 and 0. 10 FBS in the primary culture and subcul-ture respectively.

Significance of CD133 as a cancer stem cell markers focusing on the tumorigenicity of pancreatic cancer cell lines / 대한외과학회지

PURPOSE: The cancer stem cell hypothesis states that the capacity of a cancer to grow and propagate is dependent on a small subset of cells. To determine the significances of the cancer stem cell markers CD133, CD44, and CD24 using a comparative analysis with a focus on tumorigenicity. METHODS: Four pancreatic cancer cell lines, Capan-1, Mia-PACA-2, Panc-1, and SNU-410 were analyzed for the expressions of CD133, CD44, and CD24 by flow cytometry. The tumorigenicity was compared using tumor volumes and numbers of tumors formed/numbers of injection in nonobese diabetic severe combined deficiency mice. Fluorescence-activated cell sorting (FACS) analysis was used to confirm that xenograft explants originated from human pancreatic cancer cells. RESULTS: CD133 was positive in only Capan-1, CD44 positive in all, CD24 partially positive in Panc-1. After injecting 2 x 10(6) cells, all mice administered Capan-1 or Mia-Paca-2 developed tumors, 3 of 5 administered Panc-1 developed tumors, but no mouse administered SNU-410 developed any tumors. The volumes of Capan-1 tumors were seven times larger than those of Mia-Paca-2 tumors. When 2 x 10(5) or 2 x 10(4) of Capan-1 or Mia-Paca-2 was injected, tumors developed in all Capan-1 treated mice, but not in Mia-Paca-2 treated mice. Furthermore, xenograft explants of Capan-1 expressed CD133+CD44+ and Capan-1 injected mice developed lung metastasis. FACS analysis showed that xenograft explants originated from human pancreatic cancer cell lines. CONCLUSION: CD133 positive cells have higher tumorigenic and metastatic potential than CD44 and CD24 positive cells, which suggests that CD133 might be a meaningful cell surface marker of pancreatic cancer stem cells.

The clinical significance of the detection of cell surface marker and chromosome in leukemia / 重庆医科大学学报

Objective:To investigate the clinical significance of the detection of cell surface marker and chromosome in diagnosis,treatment and prognosis of leukemia. Methods:Morphologic typing and immunophenotyping were carried out in 22 untreated patients with leukemia respectively with FAB criteria and monoclonal antibodies(McAb) which were used to label cell surface marker. Chromosomal specimens were prepared by 48-hour culture method. Chromosome was analysed by G-banding technique. Results:All the 22 cases were diagnosed as leukemia by combined detection of morphology and immunology. Among them there were 8 acute lymphocytic leukemia(ALL) in which 2 cases expressed myeloid lineage-associated antigens(My+ ALL),9 acute meylocytic leukemia(AML) and 3 chronic lymphocytic leukemia(CLL). In addition,2 cases were diagnosed as acute undifferentiated leukemia(AUL). Chromosomal karyotype analysis was carried out in 17 cases,among which chromosomal abnormalities were found in 11 cases in which 2 cases with complex chromosomal deformity had non-remission(NR). With the exception of the 2 cases aforementioned,others had complete remission(CR). Conclusion:The proper detection of cell surface marker and chromosome on limited condition is helpful for the diagnosis of leukemia,which would provide an experimental evidence for personalized treatment and prognosis.

Implantation study on the potential of murine epidermal stem cell differentiation / 中国病理生理杂志

AIM: To investigate the potential of murine epidermal stem cell (ESC) differentiation after seeded in a biodegradable carrier and implanted subcutaneously into syngeneic recipient mice. METHODS: ES cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a polyglycolic acid (PGA) net containing collagen gel, functioning as a cell carrier, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. RESULTS: The ESCs kept alive in the implant when observed under a fluorescent microscopy 3 weeks or longer after implantation, and could differentiate into hair follicle - like structure,glandular structure, and gave rise to additional structures displaying features resembling native dermis. No apparent rejection or severe side effects were observed at least 10 weeks post- implantation. CONCLUSION: It is feasible to use these ESCs as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.

Comparison of surface marker of monocyte-derived dendritic cells between cord blood and adult peripheral blood / 上海第二医科大学学报

0.05).The percentage of CD40 positive cells in CBMC-derived DC was lower than that in PBMC-derived DC[(34.80?7.77)% vs(54.37?9.57)%,P