Immortalized Human Microglia

Reference: P10354-IM

http://www.innoprot.com

P10354-IM - 1 million cells Price: 1,950.00 €

Reference for the use of the cells (same as from the Canadian company)

http://www.ncbi.nlm.nih.gov/pubmed/25561230

Hi Fulvio,

I found Prof. Kim Seung contact information: Professor (Emeritus) Division o Neurology, Dpt of Medicine, The University of British Columbia, CA

604-822-7145 [email protected]

He is the one who holds the patent on the HMO6 cell line. http://www.pharmcast.com/Patents100/Yr2004/Aug2004/082404/6780641_Microglia082404.htm

Good luck!

Hi Roland!! Thanks a lot!

Hi Roland...just a question...I'm trying to contact prof Seung for a month...and yet he have not replied to my mail., in which I was just asking to make an arrangment (also paying) for the cells...He's not answering my mail, I tried also to contact his coworker and they told me to contact him....What you think ? What could be the problem? If he keeps not answering me, I'll contact PLOS one paper editor and tell him the whole story..there's a policy about distribuiting material....

Dear Aleksandra ,

I do not have that Human Microglia cell line.

My lab-mates using Human U-87 glial (glioblastoma) cell line. please check its specification on ATCC wesite. please find the link below. Pleese confirm its properties. thank you.

http://www.atcc.org/Products/All/HTB-14.aspx

Hi Aleksandra. At last prof. Seung Kim answered...by saying ""I regret to

inform you that the HMO6 cell line is not for general distribution to

other investigators at present time." And at my request of an explanation he did not reply. So I'm contacting now the editors of different journals to ask them to contact prof Kim and to ask to share those cells. If not, he is going to face retraction of all the papers published with these cells... I cannot understand why this is a problem, I'm quite angry!

@Sagar: the U-87 cells are from astrocytic lineage, I've been working with them quite a lot and they are almost astrocytes....me and Aleksandra (as far I understand) need human, microglial, cell line...and the only pure line seems to be this damn HMO6....

Has anyone had any experience with either the C13-nj cell line or the abm SV40? I need a decent mouse line so will probably buy in the BV-2 but am still looking for a good human option. Seems like Prof. Seung is being extremely unhelpful!

You know the novelty? I've made another question, in RG, citing the latest paper of Dr. Kim, and it was CANCELLED by the admin of RG....There's something fishy in this story.....I have also contacted RG admins to ask for an explanation..let see what they say....

Has anyone used the SV40 immortalized microglia sold by Abm (cat #T0251)?

You are right Fulvio. I kept asking the researcher for this cell line for almost a year, until I finally got the same answer as you. Unfortunately, not all the people wants to share the material, but I didn't know about the policy of distributing material...

Aroa...the policy is in (almost) all important journals...unfortunately they do not want to enforce it (I have asked to retract the papers)..Maybe only Plos is still following this issue...

I am also interested in human microglial cells. Fulvio I saw that you can buy immortalized with the SV40 Abm. Are not good for your experiments?

@Fulvio Celsi

@Aleksandra

Have you succeeded in getting the human microglial cell line you were looking for? If so, please could you share the source? I need to try something on such cells.

I also did look for immortalised human microglia cell line (CHME3). I had contacted at the time Professor M. Tardieu with the same story. Based on my H&S report to work on BV2 murine cell line. The problem appeared simple. The BV2 cell line were infected with the J2 retrovirus, a non-pathogenic virus suppose to be restricted to mouse. But this was not case. When co-cultured with human cells the virus infected human cells. They don't know how exactly. They think that humans have a dormant virus called helper viruses helping the part J2 retrovirus to assemble again. The human microglia cell line was constructed as the BV2 with a retrovirus that should have not infected mammalian cells.

That's really kind but I've moved universities now but others on this thread may be interested!

@Yoelvis

I did reply to your email about CHME-5 cells. Just in case my email landed in your SPAM folder.

Given the outcome of previous attempts with the HMO6 cells, does anybody know what's the best source for CHME3 or CHME5? And, by the way, what's the difference between these two? Thanks!

The HMO6 cell line is commercially available but it is a big investment (many $Ks for the rights to use them, the cells were patented). No academic sharing of the cells has been done. I know this because I contacted the lawyer that handles the patent for the researcher that created the cell line.

Oh dear, that's bad...If published, it should have made available to colleagues. Do you know who commercialises the line? Thanks

Really !!!! how is research expected to progress?

You can order this cell line e.g. at QPS.

QPS can mean a lot of things, could you be precise.

http://www.qps-austria.com/pages/home.php?lang=DE

Best wishes, Tanja

Thanks Tanya.

I have checked the website, but I am afraid  the HMO6 human microglial cells appear not be hyperlinked to ordering details, is this temporary or does one need to call to place an order? I am assuming you have purchased the cell line from them before.

Regards

Felix

Hi every one,

So I have been reading all the comments and wondering if any one has got access to human microglial cells,

I have been looking for these cells and as Felix mentioned the QPS website doesn't have active link to access and purchase the cells.

Can any one explain what is the difference between HMC3 (Human Microglia Clone 3) CHME-3, CHME-5 and HMO6????

Are these all human microglial cells?

Has any one used Cryopreserved Human Microglia from 3H Biomedical AB? or Immortalized Human Microglia - SV40 from abm?

Thanks

Negin

Tanya, Felix and Negin.. I have contacted the company mentioned above...and this is the reply I have got: " I am very sorry but we are not allowed to share these cells with others. We can only provide service for fee with these cells!"

so..again these cells seems to be impossible to obtain...and so experiments done with that have to be considered not reproducible...call it science?!?

Negin..regarding your others question..personally I have not used those others cells lines, so I cannot say..let see if someone else answer this...

Hello Fulvio, 

If you are still interested in immortalized human microglia, you can see if the cells used in this paper is suitable for your experiment:  

Reiner, A et al. "Motor, Visual and Emotional Deficits in Mice after Closed-Head Mild Traumatic Brain Injury Are Alleviated by the Novel CB2 Inverse Agonist SMM-189." Int. J. Mol. Sci. 16(1):758–787 (2015). PubMed: PMC4307274.

Hi,

I have used CHME5, immortalized human microglia line, that we got from one of our collaborator.

http://www.ncbi.nlm.nih.gov/pubmed/?term=Establishment+of+human+microglial+cell+lines+after+transfection+of+primary+cultures+of+embryonic+microglia+cells+with+the+SV40+large+T+antigen.

Well the CHM5 that we got from different sources were all rat cells and didn't even have the SV40 antigen. I Suggest anyone working with CHM5 to analyze the cells through PCR specific for human cells. 

I agree with Y. Garcia-Mesa. Different labs that claim to work with CHME-5 cells seem to work with very different cell types. For example, some labs used them to study HIV-1 infection, although the original cell line could not be infected with HIV-1? I suggest to be very cautious when using these cells!

Hi Yoelvis:

Would mind in share your protocol to test the CHME-5? I got these cells recently and after read here I'm a little bit concerned.

Corinne:

Can you please send me the ref for the paper that shows that the original cells cannot be infected by HIV-1? Because microglia cells can be infected by HIV-1 

BIG Thanks

I don't think that it was published, but we got that information from the person who actually derived that cell line and gave it to us, Dr Tardieu. We never observed any infection ourselves. But I agree with you that microglial cells should be infected by HIV-1! This is the reason why we do not use this cell line.

I see - thank you very much for the information. Because he doesn't mention anything about this in his paper. And you also tried to infect them with HIV-1 and got nothing…good to know.

Thanks again

Angelica, We will publish the paper very soon. Please send me your email and I will share it with you. Best

Immortalized Human Microglia

Reference: P10354-IM

http://www.innoprot.com

P10354-IM - 1 million cells Price: 1,950.00 €

Reference for the use of the cells (same as from the Canadian company)

http://www.ncbi.nlm.nih.gov/pubmed/25561230

Bobbi- I was just wondering if you are currently working on these cells from innoprot and are they behaving similar to primary microglia?  I know there will be some differences, but I need some cell line to work the bugs out of my co-culture model and test over-expression plasmids, etc.

Hi, I was wondering if anybody has worked with the AMB SV40 Immortalized Microglial Cell Line (https://www.abmgood.com/Immortalized-Microglia-SV40-T0251.html#15) or the Innoprot Fluorescent Immortalized Human Microglial Cells

(Reference: P10354-IM-tGFP) (http://www.innoprot.com/en_productos.asp?idsf=72&id=11&idp=459)?

if anyone knows how to make cell ine I can provide the human primary microglia...

Hi Gilles, 

I am also interested in using a microglial cell line. Has anyone reached out to you about moving forward on this?

Dear everyone. By now HMC3 is commercially available at ATCC.

I am about to start my research project and I'm planning to use microglia cells.

To compare to previous literature it would be very beneficial for me to use N9 microglia from mice, but I cannot find a place to buy them commercially. Alternatively and closer to humans, may be the HMC3 cell line. However I cannot find a lot of research papers in which the HMC3 cell line was used. I would like to know if it is possible to activate this cell line into an activated (previously called "M2") state. As the cell line appears to constitutely produce IL-6 I have some concerns.. 

Is there anyone who is working with these cells at the moment and who could provide some additional information on the microglial properties of this HMC3 cell line?

I'm also very interested in information on the CB2 receptor expression on this cell line.

Thanks in advance.

Please see this thread about BV2 and N9 - N9 are RAT, BV2 are MOUSE. 

https://www.researchgate.net/post/What_is_the_difference_between_BV2_and_N9_microglial_cell_lines

That's interesting, because multiple papers that I read said in fact that N9 is from a murine background... I'll have to look in to this to make sure I suppose. Thanks for the info!

As I said sometimes ago in this chat, we were creating some microglia cell lines and they are available already. Maybe you guys want to check on my last paper. We have been sharing these cells. I'm not working in that lab anymore. However you can email David Alvarez-Carbonell. We have uninfected and HIV-infected clones and mixed population. Best,  

https://www.researchgate.net/publication/310663082_Immortalization_of_primary_microglia_a_new_platform_to_study_HIV_regulation_in_the_central_nervous_system

Article Immortalization of primary microglia: a new platform to stud...

I'll have a look for sure, thanks! I think via via I'm able to get the N9 cells after all, so at the moment I'm in good hands. Also it turns out N9 are DEFINITELY 100% murine.. So for everyone else, please read the original paper where the cells were created and you'll see..

Dear Quirjn.

It maybe silly question, but what is different the murine and rat?

You said N9 microglia is from mice and Bobbi said it comes from rat.

The murine is mouse and not rat?

I am also finding the microglia cell to see that how activated microglia work.

Thanks for your information about HMC3 in ATCC.

Now I am find how activated them. 

Sincerely

Will

I found human microglia immortalized cell line https://www.abmgood.com/Immortalized-Microglia-SV40-T0251.html.  It may be equibalent biological activities as well as that in HMO6.

 Hello all,

I have been following this thread for a while because I'm in the need for some microglial cell lines capable of sustaining HIV infection. I can't seem to find a good candidate for the same. As stated by Quirijn Verhoog above there have not been many reports of the HMC3 cell line from ATCC being infected by HIV. Again with the recent publication by Yoelvis Garcia-Mesa's group about possible contamination of some other common microglial cell lines I am unsure of the utility of using these cells for my research. Does anyone have any other reports of the cells form Innoprot or maybe ABMGoods (SV-40 Immortalized Microglial cells, both of them) if they are capable of being infected by HIV? Or even their CD4/CCR5 expression profiles, since these receptors are very important for HIV infection?

You can try the immortalized human microglia from ABM as Hideko recommended. I have worked with their products before and I had no problems with them. 

Has anybody worked with immortalized human microglia from

mmortalized Human Microglia

Reference: P10354-IM

http://www.innoprot.com

AIs it the same as the immortalized human microglia from ABM? I bought the cell lines last month from http://www.innoprot.com, but it seems the cells do not grow as expected, like they are growing very very slow with most cells suspensed in the medium, like to have some input with those cells  

Article Immortalization of primary microglia: a new platform to stud...

Please check out this paper: "Currently available transformed human microglial cell lines reported to be in use by other groups include HMC3 (Jadhav et al. 2014; Janabi et al. 1998; Janabi et al. 1995; Rawat and Spector 2016) and C13NJ (Martin et al. 2003). Both cell types are direct derivations of the original CHME-5 cells (Janabi et al. 1995), perhaps the most widely used microglial cell lines to study important aspects of HIV neuropathogenesis (Chugh et al. 2007; Jadhav et al. 2014; Janabi et al. 1998; Wires et al. 2012).

We have found to our surprise that CHME-5 are not human cells, as previously believed, but actually a transformed rat cell line. CHME-5 cells were originally created by transfecting human fetal microglia with the large T antigen of the simian virus 40 (SV40) (Janabi et al. 1995). We used CHME-5 cells to investigate HIV latency in microglia, and demonstrate cleavage of HIV proviruses by CRISPR-Cas9 (Hu et al. 2014). As we extended our studies to include CRISPR knockouts and ChIP assays, we began to obtain sequence information that suggested non-human sequences were present in the cells. Genotyping of the cells by microsatellite analysis (IDEXX BioResearch) also suggested that these cells might actually be of rat origin."

So apparently CHME-5 cells are not human cells, they had contamination from Rat cells. And HMC3 is derived from CHME-5, so I don't know if the one sold in ATCC is still of human origin...

I do not know what is the outcome of your experiments, but this cell line has been extensively used since 1995. The problem is that it was used under different denominations and this contributed to data fragmentation. Have a look at our recent article (https://rdcu.be/6hXK), in which you can also find relevant information on the HMC3 ATCC CRL3304 cells. I hope this can be helpful for your experiments.

Hi,

Could someone give an answer for the immortalized human microglia from ABM? did anyone working with them analyze the cells through PCR specific for human cells ? or checked if they are really what they told they are with another method thank you

Tanina Arab Take a look at this paper that did further characterization on ABM's immortalized human microglia. I hope this helps!

Stephen T. B. Lau thanks a lot