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标题:[讨论帖]细胞培养基础器材和试剂选择与购买

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最近也买了一点东西,说一个关于培养基的注意事项。

我都买的是gibco的DMEM/F12培养基,货号12400-024(大包装,这个货号在官网上可以查到),10个小包装的货号为12400-016(这个货号在官网上时查不到的)。

这种培养基在国内有四种等级。

第1种:原装进口,价格昂贵,840元,是真品

第2-4种都是假的,根据造假等级的高低价格由500-200元不等

购买培养基之前,我在园子里搜到这样一条信息:我最近买了GIBCO/INVITROGEN的DMEM/F12,按照说明书加了1.2g/l 碳酸氢钠,颜色呈橙黄色。用1 N NaOH滴定后呈橘红色至淡粉色。用之养细胞,全死了,这是为什么?以前也用过,按照上述配好后根本不用滴定PH(加1.2g/l 碳酸氢钠),溶液也呈橘红色至淡粉色,细胞长得很好。后来发现,外包装与以前买的一样,货号均是:12400-024,但是这次里边的小包装货号却是:12400-016。我查看了一下,好象手册里没有这个批号。而且,以前要隔日后颜色才变成深黄色(细胞长得很旺盛),这次上午刚加液时还呈淡粉色,下午就变成橙黄色了。换液后次日仍呈橙黄色,细胞就死了?

对于信息最后的解决办法是:我问过购买的公司了,他们说12400提示两个应该为同一成分的产品.但是12400-024可以在中国销售,而12400-016一般只在美国销售,他们答应给我换培样基!只是可惜浪费了我的血清, 细胞以及时间!

但是我咨询的最终结果却和园子里的信息不一样:

1.12400-024编号的大包装和12400-016编号的小包装是没有问题的。
2.出现细胞死亡现象的原因只有一个:培养基是假的。如果是真品的话,就不会出现园子里的问题。鉴别的办法很简单,就是查产品的有效期,但是一定要到联系gibco指定的国内代理进行查询。
3.以DMEM举例,是invitrigen公司发给我的。鉴别办法:找到包装袋上"LOT NO:1290007“(注意这个不是货号),和”EXP. 7/09“;将LOT NO:1290007报给官网后,得到1290007的效期应该是2008年6月。您购买的产品显示效期是2009年7月与记录不符。所以是假的。见下图,如果只是从包装上鉴别,是辨别不出来的。
4.如果你把LOT NO:1290007报给你购买的经销商,他们也会给你出具一个有效期的证明,但是这个证明肯定不是真的了。
5.我买的培养基还没有开始使用,等我用了以后,细胞不死,呵呵,才是最后的证据。

把这个经历发在这里,供各位战友参考!


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invitrogen细胞培养官网地址

cuturl('http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cell-Culture.html')

国内代理地址

cuturl('http://www.invitrogen.com/site/us/en/home/Global/Contact-Us.html')

刚刚知道invitrogen把gibco兼并了!
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国产二氧化碳培养箱的确质量不太好

但是我喜欢水套的,不喜欢气套的。
我们用一个水套的,一个气套的,都是上海博迅的。水套的温度不稳定,经常比设定温度高一到两度,但是温度指示很准确;气套的温度稳定,但是温度指示不准确;指示37度了,但是手伸进去感觉到很凉,绝对不到37
至于进口的质量如何,由于没有用过,所以不知道。
进口的一次性培养瓶、培养板质量都不错;国产的质量绝对比不上
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Science上面发表文章,中国国内的DMEM很多有假!
cuturl('http://www.sciencemag.org/cgi/content/full/324/5930/997?sa_campaign=Email/sntw/22-May-2009/10.1126/science.324_997')
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上文的详细内容:

Science 22 May 2009:
Vol. 324. no. 5930, p. 997
DOI: 10.1126/science.324_997
Prev | Table of Contents | Next

News of the Week
China:
Appearances Can Deceive, Even With Standard Reagents
Hao Xin*

* With reporting by Xu Zhiguo of Science News in Beijing.

Culturing immortalized human cell lines for microRNA studies had been a routine procedure in Xi Jianzhong's lab at Peking University. Then last June, something went wrong. Time after time, black spots appeared in the flasks and the cells died within a week. Xi, a biomedical engineer, spent the rest of 2008 trying to figure out why his team's experiments were failing. He finally got a tip that a cell growth medium his lab was using—Dulbecco's Modified Eagle Medium (DMEM)—might be bogus. "DMEM is so basic that we never suspected it could have problems," says Xi. Sure enough, after obtaining a fresh batch from a well-known distributor, the cell lines grew without a hitch.

China is infamous for cheap knockoffs of brand-name consumer goods. Now many scientists are discovering to their dismay that a cottage industry of faux biochemical reagents has sprung up to take advantage of China's hefty increases in R&D funding. Scientists who until recently worked overseas are especially vulnerable because they may not know which dealers to trust, Xi says. "More and more researchers are returning to China. We don't want them to waste time and money like we did," says Huang Yanyi, who like Xi is a returnee in Peking University's biomedical engineering department.

After Xi learned that many colleagues had also been victimized but kept quiet about their experiences, he contacted ScienceNet.cn, a Web site for China's scientific community. In an online survey conducted with the biweekly magazine Science News, more than half of the nearly 500 respondents reported run-ins with fake reagents, according to results posted on ScienceNet.cn on 29 April.

Exposing scams may not prevent them. Currently, no Chinese agency regulates research reagents, except those used for medical tests. A database kept by Lab-on-web (cuturl('www.bioon.com.cn')) lists more than 500 reagent dealers in China; many are small operations claiming to be distributors of foreign products. The Web site has also published the names of unscrupulous dealers.

Legitimate companies say there is little they can do about the bad apples. "It is a widespread concern throughout the life science and other industries in China," says Johnson Ho, president for Greater China at Life Technologies, based in Foster City, California. Invitrogen, a division of Life Technologies, discovered that some of its products had been counterfeited. It has created a list of authorized distributors (cuturl('www.invitrogen.com/site/us/en/home/Global/Contact-Us/RegionalContactUs/China.html')). Ho suggests customers buy only from distributors on the list.

Earlier this year, Xi confronted the Beijing-based distributor who sold his lab the false DMEM. The dealer challenged Xi to prove his case. Xi called Invitrogen's office in Shanghai, which on 20 March dispatched representative Ju Jun to Xi's lab to examine the DMEM. At first glance, the packages looked genuine, but a search in Invitrogen's database by lot number, which anyone can do on the company's Web site, revealed that one lot number did not exist, and a second had an expiration date that was different from that stated on the fake DMEM's package.

Counterfeits are not limited to Invitrogen products. In recent years, others have reported fake enzyme-linked immunosorbent assay kits and ovalbumin products. More than time or money may be at stake, says Huang. If Chinese researchers were to publish spurious results because of fake reagents, he says, "we could lose our credibility as scientists."
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关于配制培养基的容器:容量瓶的使用方法

容量瓶是目前我所知道的定容最准确的容器。那么容量瓶该如何使用呢?

经过查阅资料,得到以下事项自觉应尤为注意,和大家分享:

1.检查:是否破损及瓶塞是否漏水。

2.洗涤:坚决不能用刷子刷,否则就不准了,只能浸泡和漂洗。

3.混匀:不能用转子,得振荡。

4.保存:不能当试剂瓶长期保存溶液。

5.干燥:容量瓶不得在烤箱内烤干,也不能在电炉等热源上加热。干燥的方法是:将容量瓶洗净后,再用无水乙醇等有机溶剂润洗后晾干,或用电吹风的冷风吹干。

尤其是第2条和第5条,一定要注意。不知道的要投票哦,呵呵

具体的细节请参考:医用化学实验技术导论,闫福林主编,成都科技大学出版社。

容量瓶的主要用途是配置标准溶液用,比如调节ph计的时候,我想用来配置培养基应该更准确一点,一个博士跟我说,如果你配培养基足够标准的话,那么配好的培养基的ph值根本不用调,直接就是准的。

等我试好了,我再分享经验。可怜我已经把两个容量瓶烤干了,3天3液才烤干的啊
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为了挽救我已经烤干的容量瓶,我在dxy上进行查找,结果出乎意料之外:
都知道不能烤干,但是大部分还是烤干的,一般说50度以下都不要紧,但是也有105度,200多度的,更多的用的是超声。
我昨天又泡了几个,刚刚冲洗了一下,用无水乙醇润洗后,放在37度烤箱里了。一会拿出来去泡酸。
呵呵
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为了挽救我已经烤干的容量瓶,我在dxy上进行查找,结果出乎意料之外:
都知道不能烤干,但是大部分还是烤干的,一般说50度以下都不要紧,但是也有105度,200多度的,更多的用的是超声。
我昨天又泡了几个,刚刚冲洗了一下,用无水乙醇润洗后,放在37度烤箱里了。一会拿出来去泡酸。
呵呵
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上文的详细内容:

Science 22 May 2009:
Vol. 324. no. 5930, p. 997
DOI: 10.1126/science.324_997
Prev | Table of Contents | Next

News of the Week
China:
Appearances Can Deceive, Even With Standard Reagents
Hao Xin*

* With reporting by Xu Zhiguo of Science News in Beijing.

Culturing immortalized human cell lines for microRNA studies had been a routine procedure in Xi Jianzhong's lab at Peking University. Then last June, something went wrong. Time after time, black spots appeared in the flasks and the cells died within a week. Xi, a biomedical engineer, spent the rest of 2008 trying to figure out why his team's experiments were failing. He finally got a tip that a cell growth medium his lab was using—Dulbecco's Modified Eagle Medium (DMEM)—might be bogus. "DMEM is so basic that we never suspected it could have problems," says Xi. Sure enough, after obtaining a fresh batch from a well-known distributor, the cell lines grew without a hitch.

China is infamous for cheap knockoffs of brand-name consumer goods. Now many scientists are discovering to their dismay that a cottage industry of faux biochemical reagents has sprung up to take advantage of China's hefty increases in R&D funding. Scientists who until recently worked overseas are especially vulnerable because they may not know which dealers to trust, Xi says. "More and more researchers are returning to China. We don't want them to waste time and money like we did," says Huang Yanyi, who like Xi is a returnee in Peking University's biomedical engineering department.

After Xi learned that many colleagues had also been victimized but kept quiet about their experiences, he contacted ScienceNet.cn, a Web site for China's scientific community. In an online survey conducted with the biweekly magazine Science News, more than half of the nearly 500 respondents reported run-ins with fake reagents, according to results posted on ScienceNet.cn on 29 April.

Exposing scams may not prevent them. Currently, no Chinese agency regulates research reagents, except those used for medical tests. A database kept by Lab-on-web (cuturl('www.bioon.com.cn')) lists more than 500 reagent dealers in China; many are small operations claiming to be distributors of foreign products. The Web site has also published the names of unscrupulous dealers.

Legitimate companies say there is little they can do about the bad apples. "It is a widespread concern throughout the life science and other industries in China," says Johnson Ho, president for Greater China at Life Technologies, based in Foster City, California. Invitrogen, a division of Life Technologies, discovered that some of its products had been counterfeited. It has created a list of authorized distributors (cuturl('www.invitrogen.com/site/us/en/home/Global/Contact-Us/RegionalContactUs/China.html')). Ho suggests customers buy only from distributors on the list.

Earlier this year, Xi confronted the Beijing-based distributor who sold his lab the false DMEM. The dealer challenged Xi to prove his case. Xi called Invitrogen's office in Shanghai, which on 20 March dispatched representative Ju Jun to Xi's lab to examine the DMEM. At first glance, the packages looked genuine, but a search in Invitrogen's database by lot number, which anyone can do on the company's Web site, revealed that one lot number did not exist, and a second had an expiration date that was different from that stated on the fake DMEM's package.

Counterfeits are not limited to Invitrogen products. In recent years, others have reported fake enzyme-linked immunosorbent assay kits and ovalbumin products. More than time or money may be at stake, says Huang. If Chinese researchers were to publish spurious results because of fake reagents, he says, "we could lose our credibility as scientists."

很严肃的事情,希望只是个案而不会真的已经这么严重~
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现在我也正在受一些黑斑的困扰(BLACK SPOT),不知道是什么东西,看了上面的新闻,心里有点惊,难道我的培养基也是假的,这可是接近两百块钱的,说是从上海运过来的,不知道是真是假。我的也是GIBCO的Dulbecco's Modified Eagle Medium (D-MEM) (1X), liquid (high glucose),Contains GlutaMAX™-I substituted on a molar equivalent basis for L-glutamine. Contains 4,500 mg/L D-glucose and 110 mg/L sodium pyruvate.
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