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紧急求助-求sh-sy5y细胞株！
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由于制冷设备问题，之前的细胞都报废了，现需要此细胞株！求助！
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塔秋多吉 由于制冷设备问题，之前的细胞都报废了，现需要此细胞株！求助！补充一下，由于春节假期，购买需要等到2月底，实在来不及！先行谢过了！坐标北京！
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北京气温太低，寄不了。。建议同城找找，帝都院所多，问题不大。
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Ucell 北京气温太低，寄不了。。建议同城找找，帝都院所多，问题不大。确实不在同城这点费劲！
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SHSY5Y细胞培养问题
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求助:养了两个多月的SHSY5Y细胞，最初用的DMEM+15%FBS，没有贴壁现象，换了进口血清后，使用DMEM+10%FBS(GIBCO)，出现贴壁，但是始终只有些星星点点，2天换一次培养液，经过一周多的时间依旧没有任何形态变化，着急毕业，希望大神指点。
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甲醛 编辑于
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估计楼主看到楼上的各种答案都挺难过的，因为除了细胞不好和试剂质量不过关以外的，你都已经很小心了。作为海外的科研狗，在试剂和细胞都没有问题的情况下，我养过SH-SY5Y，BV-2和A172，都成功存活了。用的试剂和你的一样，除了多了Pen Strep抗生素（但如果没有污染，应该不太影响）。所以想分享一下我的经验，希望能对你有所帮助。首先说说SH-SY5Y，的确，很好养，passage和种板都不在话下，但前提是你种在flask里面的cell confluence达到80%以上。在这个之前的确非常痛苦，一开始需要复苏挺多的冻存细胞的，我复苏过1*10^6的P3 SH-SY5Y细胞，种在一个t175 flask里面，四五周过去了，都没长起来，像你说的，零星贴壁。所以后来我砸了3*10^6的细胞，差不多花了三周左右长到80%的confluence，后面操作起来就没有问题了。我能明白你要来的细胞总不希望第一次就用光，但种SH-SY5Y的确需要一些耐心。所以建议你不要把零星贴壁的细胞丢掉，再给点耐心等等看看。如果你有多的冻存细胞，可以试试种到小的flask或者dish里面，种的密度提高一些。另外，SH-SY5Y是会出现dendrite-like structure，但除非你用retionic acid去刺激分化，不然这种结构不会特别显著。我个人觉得细胞好养程度排个名的话，BV-2 & A172 & SH-SY5Y。只要消化，medium正确，种flask的程序没有错，即使用非常少量的BV-2细胞也能很快在一周内长满一个t175 flask。然后接下来要担心的另一个问题就是，这个细胞长得实在太快了。消化，种板隔夜，再换液去血清starve之后，最好就要立刻处理了，不然再过几天medium黄掉，细胞就飘起来了。所以我们实验室并不喜欢处理BV-2超过2天，因为那时候细胞状态就不一定能有保证了。毕业着急完全能理解，我以前也一天跑过16块western的胶，但结果有时候还不如一天跑4块胶来得理想。做实验还真的是着急不得。宁可慢一点稳一点，也比不断重复失败的实验强。希望这些内容可以给你一些帮助，祝你顺利毕业！
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yucci 编辑于
一个好好少年 fly冷暖 我最初我采用的是国产血清，后来没有贴壁现象，又上网查了资料，换成了GIBCO，之后有贴壁现象，就是只有星点，没有任何其他形态变化，现在在寻找原因。养细胞就是这样子，抓紧找原因吧，刚开始我也养不好这个细胞，后来一个师姐养得好我去看了一下操作，我少了消化一步，后来就好了，现在又遇上别的难养的BV2，也是折腾了好几个月了，时好时坏，哎，加油吧
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我有这个细胞，养的还可以，用的是DMEM+10%FBS+1%双抗，细胞两天传一次，血清是GIBCO澳洲胎牛血清
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关于丁香园SHSY5Y_百度百科
声明：百科词条人人可编辑，词条创建和修改均免费，绝不存在官方及代理商付费代编，请勿上当受骗。
SHSY5Y 细胞衍生于人的神经母细胞瘤细胞系SK2N 2SH
它具有,表达儿茶酚胺能神经元特有的酪氨酸羟化酶、多巴胺2B2羟化酶和多巴胺转运体, 该细胞系被广泛运用于PD 发病机制的研究。14被浏览525分享邀请回答0添加评论分享收藏感谢收起关注今日：92 | 主题：531655
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【求助】SH-SY5Y 可以直接用来模拟AD细胞模型吗
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这个帖子发布于4年零163天前，其中的信息可能已发生改变或有所发展。
问题已解决悬赏丁当:2
之前的PC12细胞给养坏了，手里还有SH-SY5Y，这个细胞可以直接用来充当神经细胞吗？看的之前的文献大多是转染一些AD致病基因如APP或BACEmRNA之类的，可以直接拿来用吗？还有我的PC12，是未分化的，如果复活的话，必须要加神经生长因子处理使之分化吗？未分化的可以看它的突触之类的吗？
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Cell culture and differentiationSH-SY5Y cells (ECACC; Sigma Aldrich, St. Louis, MO, USA) were cultured in MEMGlutamax medium supplemented with 10% fetal calf serum (FCS; PAA Laboratories, Pasching, Austria), 50 U/ml penicillin, 50 ?g/ml streptomycin, and 2 mM L-glutamine (all from Gibco, Paisley, UK), in 37 ?C, humidified air with 5% CO2. The medium was changed twice a week and cells were split at about 80% confluence. Two different batches of cells were used for experiments, and cells were never cultivated beyond passage 25. Cells to be treated with RA or BDNF were pre-differentiated with RA (10 μM; Sigma Aldrich) for 1 week. Cells were harvested by trypsination and the cell suspension was mixed 1:1 with prechilled ECM gel (Sigma Aldrich) and seeded in pre-cooled 24-well plates (BD Falcon, Franklin Lakes, NJ, USA) at the density of 200'000 cells/well. The gel was left to set at 37 °C before the culture medium (supplemented with RA when applicable) was added, and the cells were allowed to settle in the ECM gel for two days prior to stimulation. Subsequently, different combinations of RA (10 μM), BDNF (50 ng/ Peprotech, Rocky Hill, NJ, USA), neuregulin β1 (NRG; 10 ng/ml), NGF (10 ng/ml, R&D systems, Minneapolis, MN, USA), vitamin D3 (VitD3; 24 nM), and cholesterol (10 ?g/ Sigma Aldrich) were added to the culture medium (see table 1 for details). The culture medium was changed every 3-4 days, supplemented with fresh additives. Serum-free culture medium was used whenever BDNF was included [12]. Since differentiation of SH-SY5Y cells with RA is the most commonly used regime, this treatment was used as reference in all experiments apart from the tau mRNA 6
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about PC12 cell protocolsDifferentiation:--PC12 cells should be 50-70% confluent for differentiation.--Differentiate in DMEM-Hi with 15% FBS supplemented with 50 ng/ml NGF.--Change media to new NGF-containing media every 2-3 days, as above--Differentiation is complete after 5-7 days%20protocols.htmThe rat pheochromocytoma PC12 cell line has become a commonly employed model system for studies of neuronal development. This clonal cell line, originally developed by Greene and Tischler in 1976, was derived from a transplantable pheochromocytoma (adrenal gland) tumor discovered in a rat that had been irradiated. For more than 20 years now, researchers have been using secondary lines of the original clonal cells to produce breakthrough results in neurobiological research. When cultured in a medium containing animal blood serum, PC12 cells adopt a round and phase bright morphology and proliferate to high density. Under these conditions, PC12 cells display many of the properties associated with immature neural crest cells that are destined to become adrenal gland chromaffin cells or sympathetic neurons. However, when grown on collagen coated plates, which serve as an adhesion substrate, these cells can cease proliferating and undergo differentiation in the presence of specific trophic substances, hormones, glucocorticoids, or other chemicals. In this coming week's experiments, you will be studying the effects of three drugs on the PC12 cell line: nerve growth factor (NGF), dexamethasone, and forskolin. You will also be given the opportunity to add a fourth substance to a subset of your cells to observe the effects. During the course of this experiment you will monitor cell growth and cell morphology and then determine acetylcholinesterase activity in these cells. What follows are more specific details about each of these drugs so that you can begin to hypothesize how each drug might effect the PC12 cells.NGF was first characterized by Hamburger and Levi-Montalcini (1949) as a diffusible agent which strongly promotes fiber outgrowth of sensory neurons in chick embryos. NGF is thought to be provided by peripheral tissues for the guidance and sustenance of outgrowing embryonic sympathetic and sensory neurons. In vivo, NGF may be involved in fetal development and nerve regeneration. Cellular receptors for NGF have been located in a variety of cell lines, including PC12 cells, and tissues, including cholinergic neurons of the brain and Schwann cells of damaged nerve axons. However, the signal transduction mechanism of the receptor has not yet been identified. The NGF you will be experimenting with is NGF-7S, obtained from mouse submaxillary salivary glands). NGF-7S is a 130 kD protein composed of 5 noncovalently linked subunits (2 alpha, 1 beta, 2 gamma).
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可以用SH-TY5Y细胞株，我们使用全反式维甲酸诱导分化的人神经母细胞瘤SH-SY5Y，分化的SH-SY5Y细胞表现出人神经元的特性，是一种理想的细胞模型。该细胞分化后具有神经元样的形态，表达神经元特异性的标志物，比如神经丝蛋白（neurofilament protein，NFP），神经元特异性烯醇化酶（neuron-specific enolase，NSE）和生长相关蛋白43（growthassociated protein 43，GAP-43），以及神经元极性标志物tau蛋白和微管相关蛋白2 （microtubule-associated protein2，MAP2）。
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