贴壁细胞

贴壁细胞不添加CO2培养初步探索

Preliminary study on culturing adherent cells without adding CO_2

MTT实验观察贴壁细胞的增殖活性。

MTT assay was used to study the activity of proliferation .

48h后贴壁细胞部分为成纤维样细胞；

48 hours later , part of the adherent cells became fibroblasts ;

人脾贴壁细胞中TNFα转换酶cDNA的克隆以及LPS对其表达的影响

The Cloning of TNF - α Converting Enzyme cDNA and Effects of LPS on Its mRNA Expression in Human Spleen Adhension Cells

结果：培养第3d开始出现梭形贴壁细胞，约第7d出现团状聚集细胞集落；

Results After 3 days culture , attached spindle-like cells Appeared and a number of cell clusters appeared within 7 days .

结果：诱导培养5～7天后的非贴壁细胞具有典型的DCs形态，无明显的增殖趋势；

Results : After5to7days ' culture , the non-adherent cells had the typical morphology of dendritic cells , which had no significant proliferation trends .

贴壁细胞均匀分布，3～5d增殖迅速。

Adherent cells distributed evenly and proliferated quickly within 3 to 5 days .

结果：1，分离培养的贴壁细胞（P3）具有典型的成纤维细胞样形态和BMSCs细胞表面标志。

Results : 1 , The cultured BMSCs in P3 possess typical fibroblast-like morphology and cell surface antigen of BMSCs .

利用ABC法对胎儿骨髓细胞长期培养贴壁细胞层的6种凝集素受体的分布及动态变化进行定性分析。

The localizations and changes of6 lectin receptors of the adherent layer cell in long-term bone marrow cultures of human fetus were studied by ABC method .

长期培养的急性粒细胞白血病患者骨髓贴壁细胞中src原癌基因的高水平表达

High Expression Level of src Oncogene in the Adherent Layer Cells in Long-Term Bone Marrow Culture from Patients with Acute Myeloid Leukemia

结果1.全骨髓培养法获得的贴壁细胞克隆数明显多于Percoll密度梯度离心法（P＜0.05）。

Results : 1 . The number of adhesive cell clone by whole bone marrow culture is more than by Percoll density gradient centrifugation ( P 0.05 ) .

第2章采用（31）~pNMR方法考察了贴壁细胞及血小板的含磷代谢物以及细胞内pH。

Chapter two : ( 31 ) ~ P NMR was used in the research of some cells and platelets samples for the determination of pH and the study of phosphorous metabolism .

原代培养5～7d后贴壁细胞呈梭形成纤维样细胞和圆形巨核细胞。

There were two types of adherent cells after 5 to 7 days primary culture , fibroblast-like cells and round megakaryocyte .

在不同时间点收集贴壁细胞，以RT-PCR法检测细胞因子/受体，吸取细胞培养上清以ELISA法定量检测TNF-α。

Culture cells were harvested at different time points of incubation for the detection of cytokine / receptor mRNA expression by means of RT-PCR and culture supernatants were assayed for TNF-a production by ELISA .

在M-CSF、IL-3培养基中培养贴壁细胞6天后，可观察到细胞呈圆形，集落成群生长，细胞趋向融合。

Adherent cells cultured in the presence of M-CSF and IL-3 for 6 days acquired a colony morphology , cells became confluent .

结果脐血贴壁细胞呈长梭形、针形或三角形，增殖周期G0/G1为85.12%，倍增时间为48h。

Results Cord blood adherent cells display fibroblast-like or spindle shape , and their reproductive cycle : G_0 / G_1was 85.12 % , the doubling time was 48 hours .

比较三种分离方法分离的MNC数、第三天贴壁细胞数、原代培养时间、第三代细胞数。

MNC number of adherent cells on the third day , the primary culture time , the third generation of cells with three methods of separation wre compared .

脐血CD34+细胞经SCF和TPO刺激，扩增潜能较高，体外扩增后可以获得少量贴壁细胞，传代后不再贴壁生长。

CD34 + cells proliferation is high by SCF and TPO activated . CD34 + cells were able to generate very few adherent cells and not proliferate after passage .

超过85%体外培养的贴壁细胞都特异性地摄取了Dil-Ac-LDL和FITC-UEA-1。

And more than 80 % EPCs could take up Dil-Ac-LDL and FITC-UEA-1 .

采用β-甘油磷酸钠、维生素C、地塞米松联合诱导，使胎盘贴壁细胞分化为成骨细胞，诱导后10d行茜素红染色；

Cells of 3 passages were induced to differentiate into osteoblasts with vitamin C , dexamethasone and sodium β - glycerophosphate , and the induced cells were observed in the change of matrix mineralization with alizarin red staining ;

骨髓MNC贴壁细胞形态均一，呈典型的成纤维细胞样，而脐血贴壁细胞未获得典型的成纤维细胞样细胞，混有大量的圆形细胞。

The form of bone marrow MNC adherent cells was uniform and showed the shape of fibroblast , which was not found in UCB adherent cells but mixed with amounts of round shape cells .

收集悬浮和疏松贴壁细胞，然后用免疫磁珠分选纯化，获得iDC，并用流式细胞术鉴定其纯度。

On day 7 , the non-adherent cells and loosely adherent cells were harvested , the cells labeled with bead-conjugated anti-CD11c mAb were purified and then the purity of iDCs populations was identified by flow cytometry .

而对K562和SGC7901细胞杀伤活性无明显差异。结论肿瘤周围转移引流淋巴结贴壁细胞在体外能定向诱导扩增出大量DC：DC经自身肿瘤抗原冲击后能明显提高CIK细胞对自身肿瘤细胞的杀伤活性。

Conclusion The wall cells of transfer lymph nodes around the tumor can induce and amplify multitude Cs , and they can increase the killing activity of the tumor cells once lashed by the antigens .

贴壁细胞在成骨诱导剂作用下，碱性磷酸酶（ALP）水平升高，ALP钙钴染色呈阳性反应，钙结节茜素红染色反应阳性。

It was clarified that the cultured cells were originated from stem cells . The adherent cultured cells increased the activity of ALP , ALP staining reaction and calcium nodus alizarin red staining reaction were positive with the osteogenic inducer .

α-晶体蛋白干预大鼠RGCs过氧化损伤作用实验研究给药48h后，分别取培养上清及贴壁细胞从以下四方面进行指标测定：1.观察培养细胞的过氧化损伤；

Experimental study of intervention effect of α - crystallin on superoxide damage RGCs exposed to H_2O_2 24 hours , the medium and the cell were assayed respectively as follow : 1.Observing the superoxide damage in different groups ;

而第2个48h贴壁细胞CD44+、CD34+分别占53.3%、4.6%。

There were 47.2 % CD_ ( 44 ) ~ + cells and 6.7 % CD_ ( 34 ) ~ + within the first 48 hours adherent cells , while 53.3 % and 4.6 % in the second 48 hours adherent cells .

结果1、MSCs可在体外分离扩增，其表面抗原CD44阳性而CD45阴性贴壁细胞呈长梭形，为单个或几个细胞的克隆，细胞形态均一。

MSCs may separate and proliferate in vitro , were positive for CD44 and negative for CD45 ( Fig1 , 2 , 3 , 4 ) . The cells stuck the wall were spindle-shaped , single or a few cells clones and uniform .

结果：在培养初期可见大量悬浮细胞，48h后可见有散在纺锤形或梭形贴壁细胞；第4代后细胞呈形态均一的纺锤形或梭形，分布均匀。

Results A large number of suspension cells could be seen in culture initial stage , scattered fusiform-shaped or spindle-shaped adherent cells were watched after 48h ; After the 4th generation of cells appeared uniform configuration of fusiform shape and spindle-shape and distribute equally .

目的探讨人胎盘贴壁细胞（hPDAC）的分离培养及造血相关因子的表达。

Objective To isolate and culture human placenta derived adherent cells ( hPDAC ) and assay their hematopoietic growth factor expression .

人原代贴壁细胞BMSC标志物Stro-1的阳性率为（91.4±8.3）%，小鼠为（83.5±6.2）%。

The positive rate of human adherent bone marrow-derived cells for Stro-1 : BMSC marker was ( 91.4 ± 8.3 ) % , and that of mouse adherent cells was ( 83.5 ± 6.2 ) % .