Tissue culture for Primary cell isolation are often jeopardized by the outgrowth of contaminating fibroblasts. As fibroblasts usually grow at a much faster rate compared to the target cell population, their expansion has to be tightly controlled to end up with a culture as pure as possible.
Any solutions in the literature?
A search reveals that there are numerous published efforts to inhibit fibroblast outgrowth and this process appears to be tissue specific. For instance, the addition of melatonin to melanocyte preparations potently inhibits the proliferation of fibroblasts that were passed along in the isolation procedure from whole human skin. Another example is that of adenosine and its inhibition of collagen and protein synthesis in cardiac fibroblasts. Specific antibodies can act as inhibitors of fibroblasts growth and proliferation. A DNA topoisomerase inhibitor (camptothecin) can be included in culture medium to eliminate non-neuronal cell proliferation in neuron-type primary cell isolation. In addition, it has been reported that D-valine is a selective agent for rat epididymal epithelial cells, but not for rabbit epithelial cells, and that cytosine arabinoside is a simple and effective means to control the proliferation of fibroblast-like cells in both rat and rabbit epididymal cell cultures.
Together, a tissue-specific optimal fibroblast inhibitory system can be developed to improve the performance of primary cell isolation.
Remove unwanted cells with FibrOut™
The fibroblast growth control system, FibrOut™ developed by CHI Scientific is a proprietary combination of several biochemical compounds, antibodies, or specialty reagents, which prevent overgrowth of fibroblastic or contaminating cells, effectively increasing the yield of target cells. Each fibroblast growth inhibitory system is customized to be tissue-specific and cell-specific.
Each FibrOut™ system may contain one or more of the following chemicals (1) Trypsin; (2) Collagenase; (3) D-valine; (4) Cis-OH-proline; (5) Ethylmercurithiosalicylate; (6) Phenobarbitone, (7) Formulated serum substitutes. Some of the systems also contain antimesodermal antibodies.
FibrOut™ is available throughout Europe from tebu-bio, get in touch if you have any questions or need to optimize your Primary cell culture, we’ll be pleased to help!
56 responses
How good is your Fibrout system in eliminating fibroblasts that get isolated along with smooth muscle cells.
Dear Chandra,
Thank you for reading our blog.
For mouse tissues, the FibrOut™ system recommended should be the Mouse FibrOut™ 3 (cat. nr 4-20505 & 4-20506 – for bone, heart and muscle) while for human samples it should be the Human FibrOut™ system 3 (cat. nr 4-21550 & 4-21551 for cell culture from Human tissues of bone, heart muscle).
I hope it helps.
Regards,
Philippe
How good is your Fibrout system in eliminating fibroblasts from mouse derived tumors. We are trying to establish a cell line from tumors, and can not seem to get rid of contaminating fibroblasts.
Dear Sonia,
The most appropriate FibrOut items for eliminating fibroblasts from mouse tissues are listed below:
Mouse FibrOut™ 1, for adipose, fat, eye, lung, mammary
Mouse FibrOut™ 10, for colon, intestines
Mouse FibrOut™ 11, for blood vessels, endothelial tissues
Mouse FibrOut™ 2, for cartilage, liver
Mouse FibrOut™ 3, for bone, heart, muscle
Mouse FibrOut™ 4, for brain, neural
In North of America, please feel free to contact CHI Scientific (help@chiscientific.com) for any further assistance.
Regards,
Philippe
What solution can I use to get rid of fibroblasts in a Mouse renal cell culture?
Dear Kasun,
Thank you for reading our blog.
For mouse renal tissues, we recommend the Mouse FibrOut™ 5. The Mouse FibrOut™ system 5 is a combination of several biochemical compounds and reagents, which prevent overgrowth of fibroblastic and contaminated cells and increase targeted cells growth during primary cell culture from Mouse tissues (ex. adrenal, kidney, pancreas, pituitary).
Best regards,
Philippe
Hi ,
What kind of FibrOut™ should I use to get rid of fibroblast in a mause lacrimal gland epithelial cell culture?
Dear Daniela,
Thank you for reading our blog.
For mouse lacrimal tissues & cells, the FibrOut™ system recommended should be the Mouse FibrOut™ 1 (cat. nr 4-21501 ; http://www.tebu-bio.com/Product/2384-21501/Mouse_FibrOut_tm_1_for_adipose_fat_eye_lung_mammary.html).
I hope it helps.
Regards,
Philippe
http://www.tebu-bio.com/Product/2384-21501/Mouse_FibrOut_tm_1_for_adipose_fat_eye_lung_mammary.html
What would you suggest for removal of fibroblasts from pancreatic tumour tissue
Dear Sam,
Thank you for your question.
I recommend to use the Human FibrOut 5 for adrenal, kidney, pancreas, and pituitary gland. It’s reference is 4-21554.
You will find more information at the following link.
I hope this helps,
Kind regards,
Isabelle
We are developing human prostate cancer cell lines. Which fibro-out should we use? Do I need the full kit or can I supplement my current complete medium with your fibro-out?
Dear Ann Marie,
Thank you very much for your question on our blog.
For prostate cells, I recommend you use the Prostate FibrOut Fibroblast inhibitory system.
Its reference is 7-15089. You will find full information at the following link.
You will be using your own medium, just diluting the FibrOut in it.
I hope this helps. Feel free to come back to me if you needed more info.
Kind regards,
Isabelle
We are developing a primary cell culture of human enteric neurons and glial cells. What would be the best fibro-out for this culture?
Dear Alix,
Thank you for reading our blog.
The best Fibr-out system for your application would be the Human FibrOut™ 4, for brain, neural (cat. nr: 4-21552 and 4-21553 – https://www.tebu-bio.com/Product/2384-21552/Human_FibrOut_tm_4_for_brain_neural.html).
Please do not hesitate to contact Catherine Evans (tebu-bio UK – Tel: +44 (0)1733 421880) for any further assistance.
Best regards,
Philippe
Dear Dr. Phillippe:
we isolated the endothelial cells from mouse lung, however, the fibroblast are crazy. Which fibrout kit is proper for these endothelial cells and how many times I should treat it? Once I split the cells, should I treat the cells with Fibrout kit again? Please e-mail me detail.
Dear Weiguo,
I would recommend you the Mouse FibrOut™ 1, for adipose, fat, eye, lung, mammary (cat. nr 4-21501).
The Mouse FibrOut™ system 1 is a combination of several biochemical compounds and reagents, which prevent overgrowth of fibroblastic and contaminated cells and increase targeted cells growth during primary cell culture from Mouse tissues of adipose, fat, eye, lung, mammary.
Each 1 ml of FibrOut™ system contains sufficient materials for making up 500 ml of culture medium to be used all over your cell culture experiment.
Regards,
Philippe
I am developping primary cultures from mouse PDX models (PDX are from breast cancers). Which kit should I use to get rid of mouse fibroblasts ?
Thank you in advance
Adeline,
Merci de nous suivre sur notre blog.
Puisque vous travailler sur un modèle PDX, le Mouse FibrOut™ 1 (ref. 4-21501), qui est compatible avec des tissus adipeux, l’œil, le poumon et les tissus mammaires, semble approprié.
Vous trouverez plus d’information sur ce kit ici: https://www.tebu-bio.com/Product/2384-21501/Mouse_FibrOut_tm_1_for_adipose_fat_eye_lung_mammary.html
Cordialement,
Philippe
Dear Dr. Philippe,
We isolated endothelial cells from human breast primary tumours. What reagent should we use to eliminate fibroblasts from these types of tissues
Dear Laurel,
Thank you for your interest in reading our blog.
The Human FibrOut™ system 1 is a combination of several biochemical compounds and reagents, which prevent overgrowth of fibroblastic and contaminated cells. It increases growth of your cell of interest during primary cell culture from Human tissues of adipose, fat, eye, lung, mammary. Therefire, I would recommend you this kit: Human FibrOut™ 1, for adipose, fat, eye, lung, mammary (cat. nr 4-21546 – https://www.tebu-bio.com/Product/2384-21546/Human_FibrOut_tm_1_for_adipose_fat_eye_lung_mammary.html).
Feel free to contact me for any further assistance.
Regards,
Philippe
Dear Philippe,
I isolate and culture endothelial cell from human synovial tissue.Which system will you suggest to use to prevent overgrowth of fibroblast cells? My email: bf995@bwh.harvard.edu, you can use my email to answer me.
Thank you
Bihua Feng
Rheumatology Department
Brigham and women’s hospital
Dear Bihua,
Thank you for reading our blog.
For isolation of endothelial cells, it is recommended to use a dedicated FibrOut formulation: Human FibrOut™ 11, for blood vessels, endothelial tissues.
I hope it helps.
Regards,
Philippe
Where can I find protocol? I am interested in using FribroOut or even checking the kit for isolation and growing of tumour cells.
we work on pancreatic endocrine tumours.
Dear Magdalena,
Thank you for reading our blog.
For tumor tissue, it is more effective working with the Fibrout versions dedicated to tumor tissue compared to the normal tissues (here pancreas).
Therefore, the fibrout cat. nr 4-21562 Human FibrOut™ 9, for tumors is recommended.
I hope it helps.
Regards,
Philippe
Dear Philippe,
I’m developing the primary tumor cell culture from human melanoma tissue. I found there’re around 30-40% of cancer associated fibroblast cells in the culture. How to remover these fibroblast cells? what kind of FibroOut can be used to get rid of fibroblast form the culture.
Thx,
Yan
Dear Yan,
Human FibrOut™ 9 has been designed especially for tumor tissues.
This one could be an option for your application.
Regards,
Philippe
Dear Philippe,
I isolated cells from mouse trachea. I am interested to isolate smooth muscle cells but i noticed that they were contaminated with fibroblast cells. May I know which system would work better to get rid of fibroblast cells?
Thanks,
Khan
Dear Khan,
For Lung tissues, the most adapted kit is the Mouse FibrOut™ 1, for adipose, fat, eye, lung, mammary.
I hope this helps.
Regards,
Philippe
Dear Philippe,
I am culturing schwann cells from adult rat major pelvic ganglia (MPG) and getting fibroblasts. is there any system developed to get rid if them beside using d-valine medium or using anti-thy1.1/rabbit complement treatment?
Thank you so much in advance!
Elena
Dear Elena,
Thank you for your question.
The FibrOut kit I could recommend would be our Rat FibrOut 4 for Rat brain cells (4-20533).
However, it has not been specifically tested for your cell type.
Let me know if you would like to try it. I would put you in contact with my colleagues based in the US.
Kind regards,
Isabelle
Dear Isabelle,
Thank you very much for the information. And yes, I would like to find out more about the kit you mentioned and would appreciate if I can get more information on how it works!
Sincerely,
Elena
Dear Elena,
I have replied by email with my colleagues contact details in the US.
Good luck with your experiments!
Kind regards,
Isabelle
Thank you so much for your help Isabelle!
Elena
Dear Philippe,/ Isabelle
I isolated endothelial cells from bovine aorta but i noticed that they were contaminated with a lot of fibroblast cells. May I know which system would work better to get rid of fibroblast cells?
Thanks,
yulianna
Dear Yulianna,
Thank you for your comment on our post.
I’m afraid we do not have a Fibrout kit suited to this tissue type.
You may wish to check our offer of Bovine primary cells for which we do supply specilized media to optimize the culture.
These media can be purchased separately.
https://www.tebu-bio.com/s/678qh0
Let me know if you need additional guidance and I’ll be happy to help.
I wish you good luck in your experiment.
Best regards,
Isabelle
Hello Philippe,/ Isabelle,
We are isolating Human Normal Melanocyte form Human eyeballs ( choroid tissue). We are seeing fibroblast cells with our primary normal melanocyte cells. What kit is best suitable to kill and purify the Normal choroidal melanocytes. Thank you
Good Morning,
Thank you for your question.
The closest I would recommend the reference 4-21546= Human FibrOut™ 1, for adipose, fat, eye, lung, mammary tissue.
It’s not specifically tested for Melanocytes but for Eye tissue. It should help though.
If you are contacting us from outside Europe, I recommend you contact our partner CHI Scientific directly at help@chiscientific.com if you wished to get a quote or to order.
I hope this helps.
Kind regards,
Isabelle
Hi Philippe/Isabelle,
I am currently isolating smooth muscle cells from mouse intestines and I am having trouble eliminating fibroblast contamination. I wanted to confirm that the appropriate FibrOut I should be using would be the “Mouse FibrOut™ 10, for colon, intestines”?
Any information would be greatly appreciated!
Thanks,
Holly
Dear Holly,
Thank you very much for your question on our post.
Indeed this is the reference you should be using.
You seem to be contacting us from Canada right?
I suggest you contact our partner CHI Scientific directly as we are their European distributor.
Best of luck with your experiments.
Kind regards,
Isabelle
Hi,
I am differentiating Schwann cells from pluripotent stam cells. And I do have fibroblast contamination. Do you have a kit which I can use to remove fibroblasts/fibroblast like cells?
best regards
Nazanin
Hello,
Thank you for your question.
You may wish to try our Human FibrOut™ 4, for brain, neural (4-21552).
However, it is designed to control fibroblast contamination when isolating cells from neural tissues.
It has not been tested when working with iPSC derived cells but may well work.
I hope this helps.
Kind regards,
Isabelle
Just for your information, we do provide Rat Schwann cells with optimized media which ensure optimal growth and fibroblast control.
Hi,
Just wondering what system would be best suited to removing fibroblasts from primary melanoma biopsies? We lose a lot of potential cell lines to fibroblast overgrowth that kills off the tumor cells we’re interested in culturing.
Thanks so much!
Hi Sameeha,
Thank you for your question on our blog.
I would say you have 2 options: either use FibrOut6 specialized in controlling fibroblasts overgrowth when isolating cells from human skin tissues, or FibrOut9 for tumor tissues.
As you are based in the US, I recommend you contact our partner CHI Scientific should you be interested to order. When it comes to reagents, tebu-bio is a European distributor.
I hope this helps.
Good luck with your experiment!
Kind regards,
Isabelle
Hi Isabelle,
I’m isolating ECFCs from human periphery blood. Which system would be suitable?
best,
Jette
Dear Jette,
Thank you for your message.
I could recommend the following system according to your question : https://www.tebu-bio.com/s/a1lfvuh9
I hope this will be helpful.
Regards,
Agnès
Hi,
I would like to know what system would be best suited to removing fibroblasts from primary buccal mucosal biopsies?
Thanking you.
Dear Rajesh,
Thank you for your comment.
I could recommend the following system according to your request : https://www.tebu-bio.com/s/6d6my3hu
I hope this will be helpful.
Regards,
Agnès
Hi,
I was wondering if FibrOut is also able to get rid of fibroblasts in co-cultures of fibroblasts with reprogrammed fibroblasts. In a way that we will have the reprogrammed fibroblasts, which are then considered iPSCs, still in there but the remaining non-reprogrammed fibroblasts (regular fibroblasts) will be removed.
Do you have a specific kit for this?
Dear Rick,
Thank you for your interesting question on our blog.
I’m afraid there is no specific kit developed for your application.
Should you have fibroblast contamination in a specific iPSC derived cell type, we may be able to find a suitable one, although the application would not have been directly validated. Indeed FibrOut kits are meant for fibroblast control for primary cells directly isolated from tissues.
I hope this helps.
Best of luck with your project.
Kind regards,
Isabelle
Hello,
I would like to know what type of FibrOut is used to remove fibroblasts from the human respiratory epithelium tissue culture?
Thank you so much.
Ps. the samples are taken from the lungs of COPD patients.
Dear John,
Thank you for your question on our blog.
4-21546 Human FibrOut™ 1, for adipose, fat, eye, lung, and mammary human tissue would be the best option.
Please contact your local distributor if you are not based in Europe should you wish to order it.
Best of luck with your project.
Kind regards,
Isabelle
Hi,
What is the best compound to use to eliminate fibroblasts from human lung organoid cultures?
Thanks
Dear Eleanor,
Thank you for your question on our blog.
The Human FibrOut™ 1 (4-21546) is recommended for lung tissue. I’m not sure whether it has been tested on organoids though.
If you are based in the US, you may wish to contact the manufacturer directly.
Let us know if you need some guidance for this.
My email address is as follows: isabelle.nobiron@tebu-bio.com.
I’ll be happy to help.
Kind regards,
Isabelle
Hi,
I am trying to isolate pancreatic cancer cells from mouse pancreatic tumors and I am having trouble eliminating fibroblast contamination. I’ve ordered FibrOut™, Mouse 5 Fibroblast Growth Inhibitors to solve this problem. My question is: does this chemical targets all fibroblasts, ie. normal and tumour associated?
Thank you.
Regards,
Lucie
Dear Lucie,
Thank you for your comment on our website.
The Mouse FibrOut 5 is actually more suited to working with normal tissues.
Mouse FibrOut™ 9, for tumors, may be a more appropriate choice.
As you seem to be based in Canada, I encourage you to contact our partner CHI Scientific directly.
Here is their email address: help@chiscientific.com
I remain available should you need further help.
Best of luck with your project.
Kind regards,
Isabelle
isabelle.nobiron@tebu-bio.com
Hi, does this product also work for chicken embryo primary cell cultures?