z*******0
发帖数: 35 | 1 我用干细胞做adipocyte的differeniation, 10天左右得到了如下图片，negative
control 没有这样的结果。
但是做Oil red staining 却没有红色的结果，
请问这是为什么？ 我图片上的vacuelo 是不是fat呢？ 有没有什么其它方法能鉴定
呢？或者我的分化时间太短？？
还有就是，我平时培养细胞的时候，有时也可以看到细胞内出现空泡状的结构，
请问那是为什么呢？
http://yfrog.com/71fatfj |
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z*******0
发帖数: 35 | 2 2楼的同志，那么我图片中的东西是什么？ fat还是空泡？
我用的是adipocyte differentiation medium, 而且control又没有这样的东西
形成，难道是空泡么？ 空泡的形成又是为什么呢？ |
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h**********8
发帖数: 650 | 3 you can pass over step 5
I saw some cells just the same as yours, they are not stained. Moreover, I
think they are different from other positive cells. It is obvious that the
adipocytes can be easily stained.
Did you induce with Drugs? Stable transfected with virus? |
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l****i
发帖数: 20439 | 4 没错儿 这就是adipocyte 给你看看我的图
染色后： |
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y****i
发帖数: 2194 | 5 你这个确实是adipocyte 而且我要是没猜错的话应该是primary cell
虽然differentiate的差了点...... |
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h**********8
发帖数: 650 | 6 Hi, Zhang2000
I finished my report. I stained some cells (without induction), however,
they are perfect Adipocytes.
How to upload pictures? |
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m******f
发帖数: 25 | 7 我用的是很多文章常用的诱导方法：原代MEF(passage<5), 长满后2天开始诱导(1uM
dexamethasone, 0.5mM Isobutylmethylxanthine (IBMX), 5ug/ml insulin.)2（或4）
天， 然后换成含insulin培养基培养6天左右，2天换次液。
可是分化效率很低，adipocyte的比例只有20%左右，一般很多文章都达到80%。
请各位帮忙诊断一下，谢谢 |
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f****p
发帖数: 18483 | 8
http://bbs.tianya.cn/post-funinfo-4023670-12.shtml
摘录如下。可惜了，好歹也算当过我徒弟了。。。
附录2：吐了大妈的简历
Haiying Zhou's Education
postdoc at University of California at Berkeley
San Francisco Bay Area
Research
Haiying Zhou's Education
The University of Texas Health Science Center at San Antonio
Ph.D., Molecular Medicine
2001 – 2008
Nankai University
B.S, Chemistry
1993 – 1997
Haiying Zhou's Publications
Dual functions of TAF7L in adipocyte differentiation
eLIFE
January 8, 2013
Authors: Haiying Zhou
The diverse tr... 阅读全帖 |
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m******g
发帖数: 69 | 9 Biochem Biophys Res Commun. 2010 Aug 6;398(4):723-9. Epub 2010 Jul 17.
Adipocyte-derived microvesicles contain RNA that is transported into
macrophages and might be secreted into blood circulation.
http://www.ncbi.nlm.nih.gov/pubmed/20621060
这篇paper说脂肪细胞分泌的microvesicle里面的RNA可以转移到macrophages里面，然
后macrophage的adiponeticn和resistin会升高。 他们的方法是，建立一个adipocyte
系3T3-L1，然后培养，然后把adipocyte的culture centrifuge, 提取出supernatant，
然后把supernatant进行ultracentrifuge，沉淀出这种microvesicle
请问这种方法是否可行，因为这步的关键是分离提纯microvesicl... 阅读全帖 |
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J*******a
发帖数: 96 | 10 This is the paper:
Human Milk and Infant Formula Can Induce in Vitro Adipocyte Differentiation
in Murine 3T3-L1 Preadipocytes
Lyle et al. Pediatric Research (1998) 44, 798–803
Not sure the journal is credible or not. Impact factor is only 2.8 or so...
That is not very good at all.
In vitro system is very artificial and very different from in
vivo human development.
Also, it's not "adult milk" but "breast milk" instead (they said it weird in
the paper "human milk"...). So what they really meant w... 阅读全帖 |
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m******c
发帖数: 2248 | 11 美国某医学杂志（IF 3分左右）的审稿机会，
这是一篇综述article，关于fat cell biology和obesity的，关键词有obesity,
adipocyte, differentiation, dedifferentiation, lipid metabolism,
adipogenesis, etc.
如果您正好是做obesity,adipocytes,lipid metabolism方向的，我可以帮忙推荐。
想被推荐审稿的同学给我站内发信吧，麻烦附上您的姓名，单位，联系方式，简要研究
方向。
明晚（7月3日晚）截至！
谢谢！Good Luck！ |
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c*******n
发帖数: 26 | 12 Diabetes. 2010 Feb;59(2):386-96. Epub 2009 Nov 23.
Differential effect of saturated and unsaturated free fatty acids on the
generation of monocyte adhesion and chemotactic factors by adipocytes:
dissociation of adipocyte hypertrophy from inflammation.
Yeop Han C, Kargi AY, Omer M, Chan CK, Wabitsch M, O'Brien KD, Wight TN,
Chait A.
Department of Medicine, University of Washington, Seattle, Washington, USA.
email
c******[email protected] |
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p*****m
发帖数: 7030 | 13 是这样 果蝇线虫都有人screen 但是有个麻烦就是两个物种都没有dedicated
adipocyte 因此能screen fat storage,不太好screen adipocyte 形成变化死掉的过程
。我就不知道这里面还有没有什么好玩的东西了，还是说大家觉得in vitro都把能做的
做得差不多了
多。 |
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w***e
发帖数: 269 | 14 显然不是lipid droplet。lipid 的refractive index比水高
，所以lipid droplet会很亮。如何亮呢？普通的显微镜不用开光源，lipid droplet也
会比背景的细胞亮很多也就很容易看到了。做过脂肪细胞分化的人就会明白，pre-
adipocytes放在显微镜上，不开光源，什么也看不到除了死细胞或者细胞残片。
adipocytes放在显微镜上，不开光源，lipid droplet看的一清二楚。这个图片的所有
结构一样亮，所以那个泡状结构里面是水不是油。还有一个就是感觉了。lipid
droplet有种饱满的质感，这个看起来像是个vacuole之类的东西。 |
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d****h
发帖数: 4291 | 15 如果是这篇文章的话
Human Milk and Infant Formula Can Induce in Vitro Adipocyte Differentiation
in Murine 3T3-L1 Preadipocytes
Lyle et al. Pediatric Research (1998) 44, 798–803
这篇文章我大概扫了一下
先说杂志Pediatric Research 影响影子不到3分，应该是NPG系列（nature系列）杂志
的最烂的期刊，可以没有之一，典型的灌水杂志
再说文章内容
lz解读的有问题，属于有点望文生义，至少对结论有夸大的成分，文章原始结论是促进
脂肪相关基因的表达，准确的说是转录水平而已
文章做的是离体细胞实验，也就是生物上常用的cell line
在培养基上长的细胞的状况和细胞在体内的状况本来就还差的很远的距离，不管是细胞
生长还是分化，离体细胞系和体内都差得很远
活体是不是能促进相关基因的表达，都还不一定能得出同样的结论
如果做药物研究，一般来说，先是细胞实验，其次才是模式动物，也就是通常说的小白
鼠，然后，n多实验之后才是上人体临... 阅读全帖 |
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d****g
发帖数: 7460 | 16 来，各路生物大拿。。。给我看看这篇文章。。。
http://www.helmberg.at/bone-metabolism.htm
１。The signal protein leptin is almost exclusively secreted by adipocytes (
an adipokine). Its long term plasma level is proportional to the size of an
individual's fat storage.
２。We do not yet understand the exact goals of leptin's bone effects. A
complete lack of leptin, anyway, results in increased bone mass.
http://www.ncbi.nlm.nih.gov/pubmed/22887758
３。 In summary, our results indicate that leptin, acting primarily through
per... 阅读全帖 |
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s**********r
发帖数: 67 | 17 Opening is available for a postdoctoral fellow at the Division of
Biochemistry, National Center for Toxicological Research/FDA, in Jefferson,
AR. The applicant is expected to examine the effects of triclosan on PPAR
pathways and endocrine systems as well as adipocyte differentiation. The
position is renewed annually for a period of 3 years. Only J1 visa will be
granted if you do not have green card/employment authority document (EAD).
You annual stipend will be ~ $58,000. The allowance for ... 阅读全帖 |
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G*******5
发帖数: 381 | 18 沾点边就行。 拜托大家帮忙，先谢谢了，随时站内联系，我就发CV给你。
有以下研究经历：
Tumor angiogenesis
DNA damage and repair
Adipose and adipocyte differentiation
Diabetes
Lipid and glucose metabolism
Epigenetics and transcription |
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q*********1
发帖数: 142 | 19 在准备RFE材料，引文很少，有一篇大酱油的文章的引文数据库还下载不了，不知道是
否有人能帮忙下载“Adipocytes in Skin Health and Disease” published in Cold
Spring Harbor Perspectives in Medicine in 2014.十分感谢。我的邮箱是
[email protected]
/* */ |
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S*********s
发帖数: 1963 | 20 包子感谢，可以发到[email protected]
/* */
Adipocyte and leptin accumulation in tumor-induced thymic involution (
International Journal Of Molecular Medicine)
Role of indoleamine 2,3-dioxygenase in health and disease (Clinical Science) |
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A*F
发帖数: 2272 | 21 有两篇文章需要reviewer
领域分别是:
1.microRNA, transcription regulation in adipocytes
2.microRNA, apoptosis, endothelial cells
每篇文章需要2-3名reviewer
**研究领域相近**的请把cv或者近期发表文章列表发到b********[email protected]
谢谢 |
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l******u
发帖数: 3169 | 22 Google了一下：
http://en.wikipedia.org/wiki/Adipocyte
After marked weight loss, the number of fat cells does not decrease (the
cells contain less fat). Fat cells swell or shrink but remain constant in
number. However, the number of fat cells may increase once existing fat
cells are sufficiently full.
但是另一段：
An average adult has 30 billion fat cells with a weight of 30 lbs or 13.5 kg
. If excess weight is gained as an adult, fat cells increase in size about
fourfold before dividing and increasing the... 阅读全帖 |
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G***a
发帖数: 27294 | 23 恰恰在这时候，日本的科学家，发现了一个惊人的结果
他们发现bisphenol A---一种经常用在有机塑料，和婴儿奶瓶上的物质，对细胞发育有
重大影响
人体发育有一种细胞叫fibroblast，它是组织纤维细胞。它的前体细胞是pre-
fibroblast。
日本的科学家发现pre-fibroblast，能在非常微量的bisphenol A刺激下，变异转化成
另一种细胞--就是脂肪细胞（ adipocytes） |
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r**z
发帖数: 1793 | 24 11年的文章，没做出什么
Effects of caffeinated and decaffeinated coffee on biological risk factors
for type 2 diabetes: a randomized controlled trial.
Wedick NM, Brennan AM, Sun Q, Hu FB, Mantzoros CS, van Dam RM.
SourceDepartment of Nutrition, Harvard School of Public Health, Boston, MA,
USA. n*****[email protected]
Abstract
BACKGROUND: Coffee consumption has been associated with a lower risk of type
2 diabetes in prospective cohort studies, but the underlying mechanisms
remain unclear. The aim of this s... 阅读全帖 |
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b****i
发帖数: 11322 | 25 既然是DOPING.....赫赫, at what Cost? from WIKI:
Game of Shadows, by reporters Mark Fainaru-Wada and Lance Williams, includes
allegations that Barry Bonds used insulin in the apparent belief that it
would increase the effectiveness of the growth hormone he was alleged to be
taking. On top of this, non-prescribed insulin is a banned drug at the
Olympics and other global competitions.
并且,看起来是BODY BUILDER比较喜欢用,ALSO FROM WIKI:
The use and abuse of exogenous insulin is claimed to be widespread amongst
the ... 阅读全帖 |
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D***a
发帖数: 516 | 26 脂肪和空泡在显微镜下很好区分，脂肪泡亮闪闪的，空泡暗暗的。 |
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h**********8
发帖数: 650 | 27 10 Days is enough.
Well, I have the same problem. Co Ask!
Some times, even the cells without the bubble (thought to be fat) can be
stained with the Oil Red.
It is tricky..... |
|
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D***a
发帖数: 516 | 29 照片上看不出来泡泡亮不亮，不大好说，觉得像是脂滴。 |
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d***y
发帖数: 15 | 30 应该是，ORO染色没做错吧，用BODIPY染下看？ |
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h**********8
发帖数: 650 | 31 But only a few part of cells can be stained.....
In the negtive control, no cell is stained..... |
|
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n*********b
发帖数: 140 | 33 They appear to be senescent cells. |
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W****7
发帖数: 426 | 34 我觉得是分化的脂肪细胞。应该没问题，你染色的protocol很成熟？我一般固定一下，
65度染色15分的。最好能说说你的实验细节。 |
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z*******0
发帖数: 35 | 35 1. stocking solution 300mg of Oil red O powder and adding to 100 ml 99%
isopropanol.
2. working solution: 3 part of Oil red ans 2 parts of DI water.
3. filter the working solution
4. Formalin fix sample. wash with DI water.
5. add 60% isopropanol and incubator for 5 mins
6. remove the Isoproponal and add Oil red for 15 mins
7. wash with tab water. |
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z*******0
发帖数: 35 | 36 楼上的同志能否发一张图片给我看看，我的问题是， 这种空泡是fat 么，
你染色之前，是否看到这种空泡，染色后空泡变成了red。
能否发一张这个空泡和fat都在一张图上的照片上来？
我用的是lonza的differentiation medium。 |
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h**********8
发帖数: 650 | 37 I have a report next Tuesday, I will try to upload a picture around
Wednesday. |
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y****i
发帖数: 2194 | 38 我可以负责任的告诉你 不知道这个是什么囊泡但肯定不是lipid droplet
这囊泡往往是你过分stress 细胞之后出现的
比如加病毒浓度太大，加药太猛之类的
10天够长了 你这个照片看起来就是没有分化
你还不确信的话可以测AP2 或者adiposin, adiponectin, 的mRNA或者蛋白 |
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z*******0
发帖数: 35 | 39 you can just upload it here,
http://www.yfrog.com/
do you have some picture that the vacuoles are negative at the same
time? I would like to compare a little with my pictures. Thank you! |
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h**********8
发帖数: 650 | 40 Hi, Zhang2000,
I posted one picture on yfrog. I was not able to take a convincing picture
to show the vacuoles.
I am going to the next round of differentiation, whenever I get the
comparision, I would post it.
Keep in touch.
http://yfrog.com/a9adipoj |
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z*******0
发帖数: 35 | 41 Thanks for your image, I will check my cells next week, we will see.
why I could not see any cells in your image?
Maybe you can use haematoxylin for cell staining? |
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c********i
发帖数: 47 | 42 我也遇到同样问题，没有染上；可能主要是isopropanol的纯度问题吧？如果有一点点
的methanol，就染不上了；我一开始用70%的isopropanol的。但是我后来用纯度很高的
稀释，就能染上了，另外注意isopropanol用梯度洗。 |
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h**********8
发帖数: 650 | 43 Hi, Zhang, I tried again with Adipogenesis, this time I did not see cells
like that
By the way, I can see the image from YFROG, try it again. |
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w***a
发帖数: 4361 | 44 试过electroporation么？
俺电转过siRNA到differentiated adipocytes，转完以后细胞挂了一些，但剩下的
knockdown effect可以到80%以上。
谢。 |
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s*********a
发帖数: 24 | 45 Nature Reviews Endocrinology 6, 71-82 (February 2010) | doi:10.1038/nrendo.
2009.264
Inflammatory lipid mediators in adipocyte function and obesity
Abishek Iyer, David P. Fairlie, Johannes B. Prins, Bruce D. Hammock &
Lindsay Brown
信箱 c****[email protected] |
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s*********a
发帖数: 24 | 46 ER Stress in Adipocytes Inhibits Insulin Signaling, Represses Lipolysis, and
Alters the Secretion of Adipokines Without Inhibiting Glucose Transport.
Xu L, Spinas GA, Niessen M.
Horm Metab Res. 2010 Jun 17. [Epub ahead of print]
信箱 c******[email protected] |
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n********k
发帖数: 2818 | 47 but to me, a bit too good/dramatic to be true...I have no doubt the
hypothesis/conclusion likely will stand but
I would think that the dramatic effects resulted from ER in these cancer
models is by far too good from what
many would expect...
Anyway, here is one of most interesting/amazing papers lately
Environmental and Genetic Activation of a Brain-Adipocyte BDNF/Leptin Axis
Causes Cancer Remission and
Inhibition
http://www.cell.com/issue?pii=S0092-8674(10)X0014-9 |
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w******a
发帖数: 1527 | 48 We borrowed the stem cells from other labs.
I was trying to induce them to differentiate into osteoblasts, adipocytes,
cardiomyocytes.
I did the q-RTPCR to test the expression of markers, however, I found that
untreated MSCs express higher markers than those treated...
What should I do next? |
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j**e
发帖数: 582 | 49 The mechanism is here published in Cell!
Basically, if one lives in a Enriched Environment like 唱着红歌or神清气爽,
his hypothalamus generates BDNF and regulates sympathoneural β-adrenergic
signaling and leptin level, which inhibits tumor.
Environmental and Genetic Activation of a Brain-Adipocyte BDNF/Leptin Axis
Causes Cancer Remission and Inhibition
http://www.cell.com/retrieve/pii/S0092867410005659 |
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