Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes

Background: Umbilical cord (UC) mesenchymal cells have the ability to differentiate into various cell types, which make them an easily obtainable source for therapeutic uses. Different approaches have been used to isolate mesenchymal stem cells (MSC). Aim: Here, we report a detailed enzymatic method where large number of cells can be efficiently isolated from the cord matrix and cryopreserved on the same day of arrival at the laboratory. Methods/Materials: Cells were successfully isolated from 12 samples, with a new procedure that uses the total length of the UC. MSC have been isolated using a detailed enzymatic method with collagenase and hyaluronidase followed by trypsin, without removing the vessels and without mincing the cord. Stem cells were measured with flow cytometry before cryopreservation and post‐thaw. Cultured cells were assessed for MSC marker expression and adherence plasticity for three passages. Multilineage differentiation was performed. Results: Nucleated cell yield was calculated at 0·95 × 10⁶/cm. MSC yield was calculated at 0·65 × 10⁶/cm of cord with flow cytometry while the mean length was 31 cm. Cultured cells expressed the mesenchymal markers CD29, CD90, CD105 and CD44. Mesenchymal marker expression remained intact over the three passages and post‐thaw. Osteogenic and adipogenic differentiation was evaluated. Conclusions: Our findings provide a fast and efficient method for mesenchymal cell isolation from Wharton's jelly using the total length of the UC. This method resulted in a large number of cells while the cells retained their mesenchymal character after thawing. This method can be easily applied, along with UC blood, for UC banking.     

2009～2011年邹城市城区凉皮、凉粉、面筋中亚硝酸盐含量调查

[目的]了解2009～2011年邹城市城区凉皮、凉粉、面筋中亚硝酸盐含量。[方法]采用自制发明定性试纸条的制备方法,现场定性后实验室按GB/T 5009.33-2008食品中亚硝酸盐与硝酸盐的测定定量。[结果]2009～2011年共检测了77份凉皮、53份面筋、20份凉粉中的亚硝酸盐含量,总的合格率为31.33%,其中2009年的合格率为24.53%;2010年的合格率为30.61%;2011年的合格率为39.58%。3年测定凉皮的亚硝酸盐含量平均值分别为36.10、30.80、19.70mg/kg,合格率分别为14.81%、19.23%、33.33%;面筋的平均值分别为25.30、22.60、9.20mg/kg,合格率分别为33.33%、41.18%、44.44%;凉粉的平均值分别为7.50、7.80、6.50mg/kg,合格率分别为3/8、3/6、3/6。凉皮平均值最高,合格率最低,面筋次之,但都逐年下降,合格率逐年升高。2009和2010年凉皮、面筋、凉粉进行比较,总的平均值差异无统计学意义(P>0.05),面筋、凉粉的合格率差异有统计学意义(P<0.05)。2009和2010年与2011年凉皮、面筋、...     

生物蜂王浆抑制腹水型肝癌细胞H22生长的实验研究

目的：观察生物蜂王浆对腹水型肝癌细胞H22体内生长的抑制作用。方法：以不同的生物蜂王浆即：经扶正抗癌1号方、以毒攻毒2号方、清热解毒3号方喂养的蜜蜂所分泌的蜂王浆分别灌喂荷H22肿瘤小鼠后，观察其体内抗肿瘤效果。并以普通蜂王浆和生理盐水灌喂为对照。结果：在所有生物蜂王浆灌喂组中，1号生物蜂王浆有明显抑制肿瘤生长和延长生存期的作用；并能显增加小鼠外周血白细胞数，升高血IL-2和IFN-γ含量；组织学检查显示，肿瘤细胞大部分变性坏死，肿瘤周围有大量淋巴细胞、浆细胞浸润。结论：1号生物蜂王浆具有明显抗肿瘤作用，其机制可能与抑制或杀伤肿瘤细胞，提高机体的免疫功能有关。     

鲜蜂皇浆、SOD对更年期妇女保健作用的探讨

探讨鲜蜂皇浆、SOD对更年期妇女清除体内氧自由基 ,改善更年期综合征症状的作用。 [方法 ]　将 79例更年期妇女随机分成两组 ,A组服用鲜蜂皇浆加SOD ,B组单纯服用SOD。服用前及服用后 3个月测定外周血凝固相自由基含量及网状结构的破坏程度 ,并观察临床症状的改善情况。 [结果 ]　A、B组都有明显清除体内氧自由基、恢复网状结构、改善临床症状作用。A组效果更优于B组。 [结论 ]　鲜蜂皇浆联合SOD比单味SOD在更年期妇女保健中更有临床意义。     

Chironex fleckeri (box jellyfish) envenomation: A case study

A 35-year-old male presented on the beach, having been stung on the leg by a box jellyfish, whilst swimming at Mission Beach, Qld.Appropriate diagnosis, the use of vinegar and pressure immobilisation bandaging and immediate pain relief helped to provide a positive outcome for this patient. The aim of this discussion paper is to present this case study and a review of the current and recommended assessment and initial management of such envenomation.     

Fundamental study of application of umbilical cord mesenchymal stem cells to the periodontium to aid healing after autotransplantation of teeth

After autotransplantation of teeth the healing of periodontal tissue regulates the patient's prognosis. Umbilical cord mesenchymal stem cells (UCMSC) have shown excellent pluripotent and proliferation potential. In the present study we investigated the characteristics and developmental capability of osteogenic differentiation to find out whether human UCMSC promote periodontal healing. UCMSC were obtained by primary culture and identified using flow cytometry. Flow cytometry, real-time polymerase chain reaction (PCR), Western blotting, assays of alkaline phosphatase activity, and alizarin red staining were used to assess the potential for hUCMSC to proliferate and differentiate in vitro. Both dentine and predifferentiated or undifferentiated cells were transplanted subcutaneously onto the backs of immunodeficient mice to mimic periodontal tissue healing in vivo. The result showed that hUCMSC were readily obtained, and expressed numerous mesenchymal stem cell markers. Expression of stemness markers decreased notably during osteogenic differentiation. Through investigation of different time points, we found that the osteogenic procedure could be activated and detected at day 7. In the in vivo experiments, the predifferentiated hUCMSC showed increased ability to form cementum-like deposits surrounded by fibroblast-like tissue on the surface of the dentine. In conclusion, the potential for proliferation and differentiation, and the ability to form cementum-like tissue, suggest that hUCMSC are promising candidates as a source of mesenchymal stem cell for sources of periodontal healing after autotransplantation of teeth.     

Collagen scaffolds with in situ‐grown calcium phosphate for osteogenic differentiation of Wharton's jelly and menstrual blood stem cells

The aim of this research was to investigate the osteogenic differentiation potential of non‐invasively obtained human stem cells on collagen nanocomposite scaffolds with in situ‐grown calcium phosphate crystals. The foams had 70% porosity and pore sizes varying in the range 50–200 µm. The elastic modulus and compressive strength of the calcium phosphate containing collagen scaffolds were determined to be 234.5 kPa and 127.1 kPa, respectively, prior to in vitro studies. Mesenchymal stem cells (MSCs) obtained from Wharton's jelly and menstrual blood were seeded on the collagen scaffolds and proliferation and osteogenic differentiation capacities of these cells from two different sources were compared. The cells on the composite scaffold showed the highest alkaline phosphatase activity compared to the controls, cells on tissue culture polystyrene and cells on collagen scaffolds without in situ‐formed calcium phosphate. MSCs isolated from both Wharton's jelly and menstrual blood showed a significant level of osteogenic activity, but those from Wharton's jelly performed better. In this study it was shown that collagen nanocomposite scaffolds seeded with cells obtained non‐invasively from human tissues could represent a potential construct to be used in bone tissue engineering. Copyright © 2012 John Wiley & Sons, Ltd.     

人脐带沃顿胶间充质干细胞移植对创伤性脑损伤大鼠脑损伤区微循环的影响

目的 探讨人脐带沃顿胶间充质干细胞(WJCs)对脑创伤区周围微循环的影响. 方法 72只健康SD大鼠采用随机数字表法分为4组:(1)假手术组;(2)移植对照组;(3)WJCs移植组;(4)正常组.每组18只.移植对照组和WJCs移植组采用Feeney自由落体硬脑膜外撞击法建立中度创伤性颅脑损伤模型,再分别注射0.3 mL的生理盐水和WJCs混悬液;假手术组颅骨钻孔后不损伤脑组织不移植细胞;正常组不做任何处理.分别于移植后3d及7d进行大鼠行为学评分;取移植后ld、3d、7d和2周的脑组织切片行免疫组织化学检测,检测大鼠损伤灶周围血管内皮生长因子(VEGF)及CD34的表达;实时定量PCR(RT-PCR)检测的VEGF mRNA表达;测定微血管面密度(MVD);移植后2周和4周行免疫组化观察胶质纤维酸性蛋白(GFAP)和神经元特异性烯醇化酶(NSE)蛋白表达水平. 结果 移植后7d,WJCs移植组大鼠mNSS评分明显高于移植对照组,差异有统计学意义(P＜0.05).WJCs移植组移植后3d、7d和2周损伤灶周围皮层脑组织VEGF和CD34的表达均高于移植对照组,差异有统计学意义(P＜0.05).移植后2周和4周WJCs移植组损伤区免疫组织化学染色均发现GFAP和NSE阳性细胞,其他3组均未发现.RT-PCR检测到WJCs组VEGF mRNA的表达量高于移植对照组,差异有统计学意义(P＜0.05). 结论 WJCs移植后可增加损伤区脑组织VEGF及CD34阳性表达,参与调控损伤区微血管的变化,改善损伤区的微循环,促进大鼠神经功能的恢复.     

补充蛋白果冻对术后患儿蛋白质代谢及伤口愈合的影响

目的探讨补充蛋白质对儿童术后蛋白质代谢和伤口愈合的影响。方法对手术治疗90例3—7岁患儿分为3组包伙供膳，分别供给普通膳食、高蛋白膳食和高蛋白＋蛋白果冻膳，连续3周。观察各组血清清蛋白、血红蛋白及伤口愈合情况。螭果实验组摄入蛋白质食品高于普通膳食组供给标准蛋白质35％，术后患儿的血清清蛋白和血红蛋白水平显著提高；伤口愈合率高于其他二组，但无显著差异（Pa〉0．05）。实验组体质量增长均高于其他二组（Pa〈0．05）。螭论给术后患儿补充蛋白果冻可促进肌肉蛋白质的合成，降低肌肉蛋白质的分解，促进伤口愈合，对术后机体康复有一定意义。