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L Wnt-3A细胞、L Wnt-3A细胞系、L Wnt-3

A小鼠皮下结缔组织细胞
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  • ¥1500
  • ATCC、DSMZ、ECACC、RIKEN
  • 江苏
  • CL1253
  • 2025年12月21日
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    • 详细信息
    • 询价记录
    • 文献和实验
    • 技术资料
    • 英文名

      L Wnt-3A

    • 库存

      100万

    • 供应商

      欣润生物

    • 肿瘤类型

    • 细胞类型

      细胞系

    • ATCC Number

      设计说明

    • 品系

      小鼠

    • 组织来源

      皮下结缔组织

    • 相关疾病

      正常型

    • 物种来源

      小鼠

    • 免疫类型

      不详

    • 细胞形态

      成纤维细胞样

    • 是否是肿瘤细胞

    • 器官来源

      皮肤

    • 运输方式

      新鲜或干冰

    • 年限

      100日龄

    • 生长状态

      贴壁生长

    一、细胞特性
    1. 细胞名称:L Wnt-3A细胞(小鼠皮下结缔组织细胞)
    2. 形态:成纤维细胞样,贴壁生长
    3. 含量:>1x106 /
    4. 污染:支原体、细菌、酵母和真菌检测为阴性
    5. 规格:T25瓶或者1mL冻存管包装


    二、细胞接收后的处理:
    1、贴壁细胞

    1. 收到T25方瓶细胞后,请检查是否漏液,如果漏液,请拍照片发给我们(冻存管细胞收到后直接37℃水浴复苏或直接放置于液氮中长期储存)。
    2. 请先在显微镜下确认细胞生长状态,去掉封口膜并将T25瓶置于37℃培养约2-3h
    3. 弃去T25瓶中的培养基,换用新鲜的完全培养基。
    4. 如果细胞长满(90%以上)请及时进行细胞传代。
    5. 接到细胞次日,请检查细胞是否污染,若发现污染或疑似污染,请及时与我们取得联系。

    2、悬浮细胞

    1. 收到细胞后,请检查是否漏液,如果漏液,请拍照片发给我们。
    2. 请先在显微镜下确认细胞生长状态,去掉封口膜并将15ml离心管置于37℃培养约2-3h
    3. 1200rpm离心5min,弃去15ml离心管中的培养基,细胞沉淀用新鲜的完全培养基重悬并培养。
    4. 如果细胞长满(90%以上)请及时进行细胞传代。
    5. 接到细胞次日,请检查细胞是否污染,若发现污染或疑似污染,请及时与我们取得联系。

                          
    本公司的细胞培养操作规程,供参考
    一、培养基及培养冻存条件准备:

    1. 准备H-DMEM培养基,90%;优质胎牛血清,10%
    2. 培养条件: 气相:空气,95%;二氧化碳,5% 温度:37℃,培养箱湿度为70%-80%
    3. 冻存液:90%血清,10%DMSO,现用现配。液氮储存。
    二、细胞处理:
    1)复苏细胞:将含有1mL细胞悬液的冻存管迅速放入37℃水浴中(水面要低于冻存管盖部)摇晃解冻,移入事先准备好的含有4mL培养基的15ml离心管中混合均匀。在1000RPM条件下离心4分钟,弃去上清液,加入1mL培养基后吹匀。然后将所有细胞悬液移入含有5ml培养基的培养瓶中培养过夜。第二天换液并检查细胞密度。
    2)细胞传代:如果细胞密度达80%-90%,即可进行传代培养。

       对于贴壁细胞,传代可参考以下方法:

    1. 弃去培养上清,用不含钙、镁离子的PBS润洗细胞1-2次。
    2. 2ml消化液(0.25%Trypsin-0.53mM EDTA)于培养瓶中,置于37℃培养箱中消化2-3分钟,然后在显微镜下观察细胞消化情况,若细胞大部分变圆并脱落,迅速拿回操作台,轻敲几下培养瓶后加入3ml此细胞的培养基终止消化。
    3. 轻轻吹打后吸出,移入15ml离心管中,在1200RPM条件下离心5分钟,弃去上清液,加入1mL培养液后吹匀。
    4. 移入到事先准备好的含有5ml培养基的T-25培养瓶中或含有14ml培养基的T-75培养瓶中培养。

    3)细胞冻存:待细胞生长状态良好时,可进行细胞冻存。贴壁细胞冻存时,先要消化处理并进行细胞计数。消化方法按照细胞传代方法的1-3步骤进行,最后的重悬液使用血清。悬浮细胞直接计数后离心,用血清重悬浮,加DMSO至最终浓度为10%。加入DMSO后迅速混匀,按每1ml的数量分配到冻存管中。本公司按每个冻存管细胞数目大于1X106个细胞冻存。

    注意事项:
    1. 收到冻存管细胞后,若发现干冰已挥发干净、冻存管瓶盖脱落、破损及细胞有污染,请立即与我们联系。
    2. 所有动物细胞均视为有潜在的生物危害性,必须在二级生物安全台内操作,并请注意防护,所有废液及接触过此细胞的器皿需要灭菌后方能丢弃。
    3. 细胞用途:仅供科研使用。

    发货方式:
    复苏后发货:我们复苏细胞后发货,货期一周左右,免运费。(气温较好建议复苏后发货)
    冻存发货(干冰运输):需额外增加干冰运费,选择干冰运输的我们发两管细胞,为了保证客户接种可靠性多发一管。(气温低于0℃须冻存发货)
    细胞发货采取专业的运输包装,并选择最快捷的运输方式(顺丰速运或其他空运快递)

    Wnt-3A Enhances Bone Morphogenetic Protein-2-mediated Chondrogenesis of Murine C3H10T1/2 Mesenchymal Cells

    We have recently reported the chondrogenic effect of bone morphogenetic protein-2 (BMP-2) in high density cultures of the mouse multipotent mesenchymal C3H10T1/2 cell line and have shown the functional requirement of the cell-cell adhesion molecule N-cadherin in BMP-2-induced chondrogenesis in vitro (Denker, A. E., Nicoll, S. B., and Tuan, R. S. (1995) Differentiation 59, 25-34; Haas, A. R., and Tuan, R. S. (1999) Differentiation 64, 77-89). Furthermore, BMP-2 treatment also results in an increased protein level of beta-catenin, a known N-cadherin-associated Wnt signal transducer (Fischer, L., Haas, A., and Tuan, R. S. (2001) Signal Transduction 2, 66-78), suggesting functional cross-talk between the BMP-2 and Wnt signaling pathways. We have observed previously that BMP-2 treatment up-regulates expression of Wnt-3A in high density cultures of C3H10T1/2 cells. To assess the contribution of Wnt-3A to BMP-2-mediated chondrogenesis, we have generated C3H10T1/2 cell lines overexpressing Wnt-3A and various forms of glycogen synthase kinase-3beta (GSK-3beta), an immediate

    Wnt-3a-dependent Cell Motility Involves RhoA Activation and Is Specifically Regulated by Dishevelled-2

    Wnts stimulate cell migration, although the mechanisms responsible for this effect are not fully understood. To investigate the pathways that mediate Wnt-dependent cell motility, we treated Chinese hamster ovary cells with Wnt-3a-conditioned medium and monitored changes in cell shape and movement. Wnt-3a induced cell spreading, formation of protrusive structures, reorganization of stress fibers and migration. Although Wnt-3a stabilized beta-catenin, two inhibitors of the beta-catenin/canonical pathway, Dickkopf-1 and a dominant-negative T cell factor construct, did not reduce motility. The small GTPase RhoA also was activated by Wnt-3a. In contrast to beta-catenin signaling, inhibition of Rho kinase partially blocked motility. Because Dishevelled (Dvl) proteins are effectors of both canonical and noncanonical Wnt signaling, we used immunofluorescent analysis and small interference RNA technology to evaluate the role of Dvl in cell motility. Specific knock-down of Dvl-2 expression markedly reduced Wnt-3a-dependent changes in cell shape and movement, suggesting that this Dvl isoform had a predominant role in mediating Wnt-3a-dependent motility in Chinese hamster ovary cells.

     

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    图标文献和实验
    该产品被引用文献

    Chicken intestinal epithelial cells were obtained from NEWGAINBIO company. Cells were cultured on 37℃, with 5% CO2, in the Ham’s F-12 Nutrient (DMEM/12) that contained the following supplementations: fetal bovine serum (5%), in-sulin (5 µg/mL), transferrin (5 µg/mL), selenium (5 ng/mL), epidermal growth factor (5 ng/mL) and penicillin-streptomycin (100–100 U/mL) for cell culturing (full DMEM/12). Experiments were performed with chicken intestinal epithelial cells and working solutions were prepared with plain DMEM/12 without supplementation. For the investigations, cells were seeded onto 96-well, 24-well or 6-well polystyrene cell culture plates.

     

    Primary hVICs (passage 2) were cultured to 50–60% confluence and infected with pGMLV-SV40T-puro lentivirus (NewgainBio, Wuxi, China) at a multiplicity of infection of 80 supplemented with 5 µg/mL polybrene (Sigma-Aldrich, Buchs, Switzerland).

     

    Tissue was cultured until cells became visible around the tissue, and when the fusion reached 90% (FIGURE 1A) §ask ¦lled with the prepared culturing medium was sent to the company for further immortalisation. Cell immortalisation was done for cell stability and longer-term use. Immortalised cells were cultured with 10% FBS and 1% PS in the DMEM medium.  After the cells multiplied and merged, they were routinely passed and grown ( NEWGAINBIO Inc. Wuxi, Jiangsu, China) (FIGURE 1B-C).

     

    Mouse primary cultured renal vascular ECs and VSMCs were obtained from Newgainbio company, which were tested by Factor VIII and α-smooth muscle actin (α-SMA), the marker of ECs and VSMCs. RNeasy Mini Kit was used for RNA extraction, and the above protocols were repeated.

     

    Porcine primary colon epithelial cells (Newgainbio company, Wuxi,China) were cultured in Dulbecco's Modified Eagle's Medium (Solarbio, Beijing, China) containing 10 % fetal bovine serum (BioInd, Kiryat shmona, Lsrael) at 37 ◦C and 5 % CO2 humidity.

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