Will McDonald’s Serve Stem Cell Burgers Someday?

Dr. Mark Post, head of physiology at Maastricht University, will debut the first stem cell burger the world has ever seen this Monday. Why? Dr. Post breaks down the theory and goals behind this lab-grown hamburger in his TEDTalk talk below. However, for those of you who do not have the 20 minutes to watch, we have attempted to explain his interesting theory in this post.

Dr. Post explains in the video how inefficient meat production is. He claims that pigs and cows never went through an evolution to become dinner for us, and therefore they are very inefficient. For every 15 grams of meat we eat, we have to feed these animals 100 grams of vegetable proteins, making their bioconversion rate 15%; and livestock is currently using 70% of our arable lands in the world. [Read more…]

Media, Media, Media – Cell Culture Media, of Course

When we are thirsty, nothing is better to me than an ice-cold glass of H20. Some people reach for something sweet, some like the carbonation, but us, we love the refreshing feel of ice-cold water. To each their own, right? Like all living things, cells need nourishment, and cell culture media is the stem cell culturevessel to this nourishment when cells are grown outside the body in a culture dish. Typically, media are water-based liquids that carry basic nutrients cells need to survive: salts, sugars, amino acids, proteins, etc. However, just as the water industry exploded with its various flavors of water, vitamin water, energy water, the cell culture media industry has exploded with so many different types and flavors—and that’s just for the basic cell culture media. [Read more…]

Stem Cell Scientists Once Again Create Hope, Now It’s Time to Convert

This past week, there was another instance of stem cell scientists creating hope.  A team of stem

stem cell growth factors liver

Reference: Examiner.com

cell scientists in Yokohama, Japan has grown tiny pieces of human liver tissue using stem cells. When the liver pieces were transplanted into lab mice and attached to the animal’s blood supply; they became functional and started cleaning toxins from the blood.  These findings are no doubt a major breakthrough for stem cell research across the globe.

In the field of regenerative medicine, this is the first instance of the production of a complex organ that responds to a recipient’s blood supply.  Once again, no doubt, this is a true breakthrough for stem cell research. [Read more…]

StemBeads FGF2 Trial Size Released

In response to requests from our colleagues, we have released a trial size (1 mL) of
StemBeads FGF2.  Many stem cell scientists who engage in stem cell culture have asked us to StemBeads FGF2provide a smaller size of the StemBeads FGF2.  This smaller size allows for a laboratory to test out the product at minimal cost.

StemBeads FGF2 is a revolutionary growth factor supplement that offers a more efficient way to grow FGF2 dependent stem cell cultures. Already verified in some of the top stem cell labs in the United States, this supplement delivers a steady release of growth factor into your media of choice creating a more stable environment. This allows scientists to reduce the number of times they feed their cells from daily, to every 3rd day.

The Benefits of StemBeads FGF2

With regards to stem cell growth, StemBeads FGF2 provides the following benefits:

  • Reduction of Media Changes by (Saves you time)
  • Significant Savings on Media Costs (Saves you money)
  • Better Quality Cultures by Reduction of Spontaneous Differentiation (Facilitates better cells)
  • Use of Your Favorite Media, No Change of Culture Conditions

Please note that 8 uls of StemBeads FGF2 are needed per 1 ml of growth media.

The science behind StemBeads FGF2 has been published in PLOS ONE.  Feel free to contact us should you have additional questions with regard to the science behind StemBeads FGF2, or any other questions about our products.  It is our goal to make your research as a stem cell scientist easier, more enjoyable, and extremely successful.

Click here to order a trial size of StemBeads FGF2 and start saving money, saving time, and grow better cells.

Let it Bead,

StemCultures

Information for scientists by scientists on all things related to stem cell growth including stem cell culture, culture medium, tissue culture, fibroblast growth factor (FGF2) and more.

Making Stem Cell Conferences Count: It’s More Than Showing Up

Next week is 11th annual International Society for Stem Cell Research (ISSCR) meeting. This meeting is the largest forum for stem cell and regenerative medicine professionals from around the world. The meeting format consists of over 2000 abstracts stem cell conference bostonand 150 speakers talking about their latest research, along with exhibits and vendors who provide stem cell reagents and tools. While this meeting has gotten bigger over the years, if you are in the world of stem cell research, you should be there. There are three things you should try to do when attending this meeting or any stem cell conference for that matter.

Learn. Spend some time prior to the event looking at the abstracts and make a plan of attack. In many case,  a lot of the data presented will either already have been published or be very close to it, so be sure to engage the speaker or presenter with follow-up questions. Even though you are in a public setting, it is still ok to be critical—after all, we are scientists, and that is what we do. In addition to learning about work in your own field, try to expand your mind a bit and read about work that might be outside of what you normally do. Sometimes it gives you great experimental ideas, as many of the approaches might be similar to yours, albeit in different contexts.

Find New Products. One of the most fun things to do is to cruise the conference floor to visit with all the companies selling their products. Not only will you score some cool free stuff, but you may find some of the latest and greatest stem cell research tools out there. Even better, go ask questions about products you already use if something hasn’t been working properly or as advertised. Our company, StemCultures, will be there this year at booth number 506 talking about our StemBead products, like StemBeads FGF2. We like to have fun, so we will be giving away Mardi Gras beads to wear and a chance to win tickets to an open bar Friday night. Please stop by our booth.

Connect.  This really is the best part of the meeting. Make sure you try to meet as many people as you can. Go to the events, make new friends and potential collaboration partners, and widen your web of stem cell colleagues. we meet so many people this way, and going to ISSCR every year allows us all the chance to be in the same place talking science and having fun.

Looking forward to seeing you all out there. If you have other recommendations or approaches that you use to get the most out of the stem cell conferences you attend, please reply to this post below.

If you enjoyed this post, please consider subscribing below to our monthly newsletter through which we provide stem cell scientists information on the most current research topics and tools to help them constantly improve their stem cell culture experience.


To Successful Research,
StemCultures

Information for scientists by scientists on all things related to stem cell growth including stem cell culture, culture medium, tissue culture, fibroblast growth factor (fgf2) and more.

Note: StemCultures facilitates posting on this blog, but the views and accounts expressed herein are those of the author(s) or interviewee(s) and not the views or accounts of StemCultures its officers or directors whose views and accounts may or may not be similar or identical. StemCultures, its officers and directors do not express any opinion regarding any product or service by virtue of reference to such product or service in this blog.

Lifting the Curtain: a Beginners Guide to iPS Cell Culture

This is a post by Christine Miller, Research Assistant at Harvard University, Amy Wagers Lab:

I think it is fair to say that most people who have experience with cell culture know that there is at least some degree of “black magic” that goes into getting a particular protocol to work. In my experience, I’ve found this to be especially the case with iPS/ hES cell culture. In this series of blog posts I hope to shed a little light on this “black magic,” to talk about what I’ve found works, and hopefully to generate a platform for others to share their secrets as well.

Even though iPS cell culture is a relatively new technology, there are already tons of protocols for culturing them—each with its own variations on the amounts of reagents to add to culture media, methods of passaging, ways of freezing down lines, and the list goes on. Clearly, there are countless variables to test if you want to optimize your culture strategy. In addition, however, I have found that not only are there variables in technique, there are also lots of differences in iPS lines, even when they are all reprogrammed from normal patients. These differences may be illuminated in ways like pluripotency tests, where one line may take exactly six weeks to form a clear teratoma while another line may not exhibit tumors until 10 to 12 weeks. This might not sound too surprising on paper, but when you have injected a couple different lines on the same day and six weeks down the road all your lines except for one have teratomas, it is easy to think that that last line just didn’t work. If you wait another couple weeks, you may be surprised to find a cage full of mice with teratomas. Also, differences in lines may become very obvious while trying to differentiate iPS cells down a particular lineage. Currently I have been working on driving cells down the hematpoietic lineage, and I’ve found that the culture conditions for differentiating one line are quite different from differentiating another line. Even variables as small as the line’s growth rate or passaging timing may be different. My point with all this is simply that you should be aware that these differences exist and to be open-minded if your experiences with one line do not translate 100% to those with another line.

So, moving on to the good stuff—how to deal with some of these variables. I’m going to give you the “Dummies” edition of what specifically I have found to work well culturing my cells.

iMEFs vs. Matrigel
There are two main ways to culture iPS cells: you can culture them on a feeder layer using irradiated mouse embryonic fibroblasts (iMEF), or you can culture them feeder free. Depending on your desired application, both methods have their benefits.

Here is the breakdown of what I have found using iMEFs.

iMEFs are really great if you are thawing a line you’ve never worked with before. They are reliable in culture for a good 10 days, which should give you enough time to see a couple small colonies form. Also, the iPS colonies formed on the iMEFs will be nice and uniformly shaped, so you will be able to clearly identify where your colonies are and where differentiation (if any) is occurring. However, there are a couple things that must be taken into account using iMEFs. First, you must use good-quality iMEFs. If they are not high quality, they will not provide the appropriate feeder layer and support that your iPS colonies need, resulting in failure to seed or improper seeding that leads to differentiation. You want your colonies to fit fairly snugly between the iMEFs so that they can stay contained and undifferentiated. However, if they are too snug (the iMEFs are plated too densely), the colonies will grow vertically and risk differentiating on account of not having the space to expand horizontally. I’ve used both homemade and purchased iMEFs and have found for my needs it is more cost effective to buy them. I get them from global stem (cat # CF-1 MEF), and it costs $24 for a vial of 2M cells. I plate them at 200k/well of a six-well plate and do this by splitting one iMEF vial over 10 six-well plate wells (ie: 1 and 2/3 plates). I’ve tried plating anywhere from 100K to 300k, and 300k was definitely way too much, but 100k was a bit too sparse for my iPS cells to seed well. Making sure they are evenly spread out over the plate is also really important, so be sure to do the “T” motion at least three times in the hood and then at least one more time in the incubator. My only comment about the homemade iMEFs is that, unless you are making them to share with many others (and therefore can take turns harvesting, irradiating, and preparing them), it’s a lot of work and may not necessarily save you that much money. The main downside to using iMEFs is that it’s a much more time-consuming process. In order to passage or seed iPS cells onto them, first you would need to gelatin-coat your plates, which will take a minimum of four hours to set. Then you can plate your iMEFs, but those need to sit overnight in order to plate properly. Ultimately, then, this means that preparing your plate needs to start one or two days before you want to plate your iPS cells on it.

Feeder free, on the other hand, is very quick to prepare, taking only one to two hours to set. The main products on the market right now for this are Matrigel (BD), CELLstart (Invitrogen), and Vitronectin XF (Stem Cell Technologies). I have only tired Matrigel, but from the descriptions of CELLstart and Vitronectin, they sound very similar. Another benefit of using one of these feeder-free systems is that they are quite a bit more streamlined and simple. The media usually comes as part of a kit, where you only have to add a couple of things (if anything) in. There is usually some sort of standardization with these systems allowing you to purchase not only your media but also a recommended passaging reagent and freezing reagent, which can be nice as well. My last comment about working feeder free is to make sure you are buying the hES-grade material. The first time I ordered Matrigel, I didn’t realize that there were differences in grade and purchased a non-ES cell-grade one. After about six days in culture, the Matrigel would degrade and my colonies would lift off the plate with the matrix and basically be completely destroyed.

Passaging
iPS cell culture 2
There are two main methods for passaging hES and iPS cells: using an enzyme or manually detaching the colonies. Depending on the status of your plate and colonies, one method may be more useful to you than the other. In my experience, when you have fewer than 20 colonies (per well in a six-well plate), it is much better to passage manually. This gives you much more control over what you are detaching from the plate and bringing over to your fresh plate. This should also then be passaged at a 1:1 split unless the colonies you have are pretty large. Even though there are lots of different methods and tools you can use for manual passaging, I’ve found the most effective way to do this is to just use a p200 pipette and tips. This seems to be the perfect size to allow you to score larger colonies into sections while also scraping up the smaller colonies with one scratch. I’ve tried using Pasteur pipettes with the tips curved using a Bunsen burner, but this seems to yield too blunt and irregular tips. I’ve also tried using different-sized needles to break iMEFs off the colonies and score the colonies into smaller pieces, but this often scrapes plastic off the bottom of the plate, getting pieces of plastic mixed into the colony.

If you have over 20 colonies per well, I think it is much easier to go with an enzymatic passage. If you are using a feeder layer, the quality of the colonies may be slightly worse than with manual passaging, because you are picking up the iMEFs in addition to your colonies, which can result in your colonies forming large clumps in the new plate rather than seeding nicely into the new feeder layer. For hES and iPS cell culture, the enzyme used shouldn’t break the cells into a single cell suspension (like Trypsin/EDTA); rather they should be broken into smaller clumps for optimal seeding and growth capabilities. I have used 1mg/ml collagenase type IV (Invitrogen) diluted with DMEM-F12 for passaging with a feeder layer, and Dispase (Stem Cell Technologies) also at 1mg/ml when passaging from Matrigel. Both collagenase and Dispase keep the cells in clumps. Once prepared, collagenase only stores for two weeks, so be sure to not hold on to it for any longer than that; the enzyme becomes weak and ineffective. Even if you are passaging enzymatically, I have found it very helpful to do a little colony cleaning manually beforehand. Getting rid of partially differentiated colonies, breaking up larger colonies into smaller pieces, and teasing away some iMEFS can really make a big difference in the quality of your cells.

When you are removing the cells from the plate after the enzyme treatment, a really effective method for scraping is the “car wash” method. This is done using a 5ml glass serological pipette. While tilting the culture plate slightly forward so that the media forms a pool at the bottom half of the well, you pull up the media, and while releasing the media, scrape in a zigzag pattern from the top of the well towards the bottom. By releasing the media while gently scraping, you help keep the colony-removal process gentle and the cells in bigger clumps. Once you have cleared the top half of the well, flip the plate around so that the other half is on top and repeat the process.

Antibiotic Use
Many people have very different views from mine on the use of antibiotics for hES/iPS culture. I feel very strongly about culturing antibiotic-free, and I will explain why. First of all, it allows you to have more control over the status of your cells. If there is any sort of breach in sterility, without antibiotics, you will immediately know and be able to deal with it by getting rid of the contaminated plates. You never have to live questioning if a plate is infected or not, meanwhile exposing your other plates to the potential infection. Everything is very clear; infections are obvious and therefore can be dealt with swiftly, without jeopardizing the rest of your cells. One of the few times in my iPS culture experience that I was using an antibiotic in my media, after not knowing whether a particular plate was infected, one by one all of my plates became infected and I literally lost every single culture I had. Now, I know that this is probably a pretty extreme case, but in any event, it demonstrated what can happen when antibiotics are battling a bacterial infection. Since the infection was not obvious, I continued to expose my cells to contamination unknowingly and therefore contaminated everything.

Secondly, using an antibiotic can mask mycoplasma infections. Usually, mycoplasma infections are accompanied by other infections—or rather, when the sterility of your cultures is breached, mycoplasma can also be introduced, and they are typically introduced with other infections such as bacteria. If the antibiotic successfully fights off the bacterial infection, your cells will still have the mycoplasma infection, which is typically only detected by using specific mycoplasma detection tests (by taking spent media and testing it). Last year, our iPS facility tested positive for mycoplasma. This was a total disaster. We had to throw away all cell cultures, close down the core for fumigation, and literally throw away all disposable materials in the room including reagents and media. It left us out of commission for a whole month. Not only was this an unbelievably expensive endeavor, it also made us lose valuable time, resources, and in some cases permanently lose cell lines. At the time when this happened, we were all using antibiotics in our media; since then, we have made it a room rule to not use them. Since we have been antibiotic free, we have also been mycoplasma free. Not using antibiotics also helps reinforce good practices in sterile technique, forcing you to be ultra careful with your cells and keep your surroundings very clean. This helps eliminate some variables in culturing, since you have more control over your environment and therefore over culture conditions.

Last notes
I think one of the most important things to remember with iPS cell culture is to be patient. Especially if you are just thawing cells for the first time! Even if it looks like there are no colonies, I would be willing to bet that if you keep feeding and wait, you will see at least one. Sometimes this can be a slow and frustrating process, but just keep at it and you’ll eventually get some great cultures. I was the first person in my lab to do human iPS cells work, so I truly understand how difficult it can be getting things up and running. There are a lot of helpful resources online, and as the stem cell community grows, the resources grow also. The HSCI iPS core has their protocols available online (http://www.hsci.harvard.edu/ipscore/node/8), which I have found to work well. WiCell has many helpful resources and protocols available online as well. I use their protocol for teratoma assays, and it pretty much works without fail (http://www.wicell.org/home/support/stem-cell-protocols/stem-cell-protocols.cmsx).

So, to wrap things up, if you are new to hES/iPS culture, I hope this has lifted the curtain a bit on culture techniques, hopefully helping to eliminate at least a couple variables while you get started. If you have tips of your own now (or later!), please do share! Pooling our secrets, we can help each other out and make some real scientific progress.

Christine Miller is Research Assistant at Harvard University, Joslin Diabetes Center, Amy Wagers Lab.

Note: StemCultures facilitates posting on this blog, but the views and accounts expressed herein are those of the author(s) or interviewee(s) and not the views or accounts of StemCultures its officers or directors whose views and accounts may or may not be similar or identical. StemCultures, its officers and directors do not express any opinion regarding any product or service by virtue of reference to such product or service in this blog.

Smooth Edges in Stem Cell Culture

This is a post by StemCultures CEO Jeffrey Stern PhD, MD.

When I think about the development of an organism, smooth edges, like the irresistible full folds and curves of a baby, come to mind. Throughout development, stem cells give rise first to the Smooth Edges in Stem Cell Cultureembryo, then later, through tissue-specific stem cells, to the final biological form, with naturally rounded shapes and few sharp edges. At the molecular level, I imagine similar changes—with gradual shifts in growth factors forming rolling gradients elastically linked to intrinsic gene regulation and protein expression. Over time and through evolution, the jagged peaks and shear faces of primitive organisms have been worn into smoother, more complex forms of life. As time softens sharp-edged mountain ranges into rounded, biologically complex environments, evolution shapes the sudden molecular and organismal changes that carve new pathways of development.

On the other hand, research seeks sharp, definitive, cut-and-dried explanations. Human nature seeks a clear path—with, when needed, stairs and a guide-rail—to follow. Indeed, the current revolution in stem cell science was enabled by the discovery of step-by-step protocols to expand cells without differentiation. Discoveries by pioneers such as Martin Evans, Irving Weissman, James Thomson, Benjamin Reubinoff, Rudolf Jaenisch, Shinya Yamanaka, Douglas Melton, and Austin Smith led to an understanding of the factors needed to maintain stem cells, providing the large quantities of cells needed for research and therapy. These pioneering results enable us to study stem cells and to discover just how complex the targeted biological processes are. As work progresses to refine protocols, they are beginning to evolve. Our recent work aims to mimic the more gradual changes, resembling environments encountered in normal development.

We’ve found that copying the normal developmental niche can have practical advantages, illustrated by our laboratory’s recent success using controlled levels of growth factor in stem cell cultures. Standard protocols add FGF2 daily in a step-wise fashion: suddenly via pipette, a bolus of this key growth factor is released onto the cells, causing a dramatic spike in levels. Surprisingly, half of the added FGF2 is lost in less than six hours, meaning there are extended periods of negligible FGF2 levels each day, followed by a sudden rise in concentration upon feeding. Using controlled-release to smooth these dramatic peaks and valleys results in more stem cells, increased stem markers, and decreased differentiation. The cells seem happier when these natural gradients replace sudden, dramatic changes in environment. For this discovery, like any other, it will take time to learn the full implications—but the philosophy of letting nature guide is one that we believe is worth following.

If you enjoyed this post, please consider subscribing below to our monthly newsletter through which we provide stem cell scientists information on the most current research topics and tools to help them constantly improve their stem cell culture experience.


Note: StemCultures facilitates posting on this blog, but the views and accounts expressed herein are those of the author(s) or interviewee(s) and not the views or accounts of StemCultures its officers or directors whose views and accounts may or may not be similar or identical. StemCultures, its officers and directors do not express any opinion regarding any product or service by virtue of reference to such product or service in this blog.

Dear Scientists: Change is good, we promise

FGF2 Change is Good Today, many can’t even remember how we lived before all of current technology, but we can maybe remember our eagerness to try them out when they were first introduced. Of course there is a learning curve, but once you become familiar with anything, over time, the new way becomes as easy as the old way was before. In fact, it should get easier, as that’s what change should do.

Scientists are a different breed. We say we are open to change in our day-to-day research, but are we really? Try going into a lab and asking the scientists to try a new reagent out. We estimate 75% would say “Thanks but no thanks, we’re good with what we’re doing.” Why is that? Scientists operate by repetition. We tend to do whatever everyone else does and live by the motto if it’s broke, don’t fix it. During grad school and even while a post-doc, Many can’t be bothered to change things; they just didn’t have the time. However, some learn that changing things can actually save you time and make your life better. Here are three reasons for us scientists to be a bit more open to change in our research lives.

Efficiency.  New technology allows for the creation of new products that will let you get what you normally do, just so much faster. Case in point, the PCR machine. How slow would your day be if you couldn’t run a PCR? Now that’s a bit drastic, but it shows the point. New products should make your life easier, save you some time and headache. If this is part of their sales pitch, try it, as a few more hours or even days back in your life is a lot.

Money.  Plain and simple, experiments are expensive. Once again, as a graduate student and post-doc, I wasn’t paying a lot of attention to this; I would just buy reagents and hope my boss OKed them later. Now, running my own lab, I pay close attention to the cost of reagents and materials and try to teach my lab members to do the same. After all, the funding rates are plummeting, and we need to stretch every dollar we can.

Improvements.  The ultimate reason to change to a new product is if it makes your experiments better. For example, if a typical yield of cells in a routine experiment is 30%, and a new product claims it can get that up to 75%, you should try to change. Sure 30% might work, but 75% obviously means you are doing something much better.

So if a new product will save you time and money and improve your output, changing over should be a no-brainer. A lot of new products don’t have all three of these qualities, and that makes it harder. Typically, new products might save you time and increase output but cost more than what you currently do. This might make it hard for you to change, or at least might make it hard for your boss to agree to change.

FGF2 Change is Good 2

Growing stem cells costs a lot money, and it’s a daily effort. Cell culture media has to be changed every day for some stem cell types, and even when researchers are doing this, the cells are imperfect, with a lot of room for improvement.  Knowing that scientists are hesitant to change, we wanted to fix this problem without drastically changing what scientists normally do. So, what is it we normally do? Every day, we refresh the media and a recombinant protein FGF2. FGF2 is essential for maintaining stem cells, and we found that it is incredibly unstable in culture and is the reason why stem cells tend to drift from their stem cell state. Changing the media every day is expensive, and that expense comes from the FGF2 and the cell culture media alone. One way we could have fixed this problem was to create a small molecule FGF2 mimic that would be more stable. Small molecules tend to be cheaper, and that would have achieved one of our goals, reducing cost. The problem there was that small molecules are very dirty and do not exist in nature. Asking a scientist to switch from protein to small molecule is a big change; we opted not to go this route.

The other option was to keep the protein FGF2, but to mutate it in a way that made it more stable. That would reduce the need to change the media so frequently. However, now we were asking scientist to change from a normal protein to a mutated one. Doesn’t sound like a lot, but it’s a major change, and we know as well as anyone that scientists don’t like change. So our solution? …Don’t change anything. Let’s just put the normal protein FGF2 in a formulation that will constantly release it over time. Kind of like a protein IV for cells. When we did this, we found that researchers saved time, saved money, and best of all, the cells they were growing were more stem-like. The trifecta. Best of all, we didn’t really alter anything: scientists still use the same media and protein FGF2, they just use less of it and save money.

There are many people using these StemBeads and enjoying all the benefits. However, there are hundreds who are not, probably either because they are not aware of them or because they are hesitant to change what they normally do. I get it; I’ve been there. But a product that does all of those things comes around once in a while. Give StemBeads a shot; it might significantly change your research world. After all, change is good, especially when it puts more money into your pocket and more time back into your day.

Please consider subscribing below to our monthly newsletter through which we provide stem cell scientists information on the most current research topics and tools to help them constantly improve their stem cell culture experience.

To Successful Research,

StemCultures

Information for scientists by scientists on all things related to stem cell growth including stem cell culture, culture medium, tissue culture, fibroblast growth factor (fgf2) and more.

Note: StemCultures facilitates posting on this blog, but the views and accounts expressed herein are those of the author(s) or interviewee(s) and not the views or accounts of StemCultures its officers or directors whose views and accounts may or may not be similar or identical. StemCultures, its officers and directors do not express any opinion regarding any product or service by virtue of reference to such product or service in this blog.

StemBeads FGF2 is now a Peer Reviewed, Published Science

StemBeads FGF2The science behind StemBeads FGF2, the novel product that provides researchers with a more efficient way to grow better stem cells is now published in the recent issue of PLOS ONE.  The study performed by a team of researchers at The Neural Stem Cell Institute led by myself highlights the instability of soluble FGF2 in cell culture, and how stabilization of FGF2 for an extended period of time greatly improves the quality of stem cell cultures.   Click here for a free copy of this published manuscript.  After you learn about this fantastic science, be sure to click on the StemBeads FGF2 product page for ordering information. [Read more…]

The “Ups and Downs” of FGF2 and Stem Cell Culture

FGF2 and Stem Cell Culture Ups and DownsStem cells are unique in that they, like humans, possess the ability to choose.   “To be, or not to be,” is appropriate, as stem cells must decide if they want to maintain themselves as a stem cell, or turn into another cell type.  Just like us, many factors influence this decision, and one of the major ones is the protein fibroblast growth factor 2, also known as FGF2.  FGF2 eliminates this decision for stem cells, by forcing them to maintain the stem cell state.  In vivo, stem cells exist in a highly organized niche, where factors like FGF2 are maintained at constant levels to ensure stem cells are kept in the undifferentiated state.

In stem cell culture, there is tremendous value for keeping stem cells in the undifferentiated state.  The ability of stem cell to differentiate into multiple cell types is directly proportional to their level of “Stemness”.  The more undifferentiated stem cells are, the more efficient they will be in generating progeny.  In cell culture, scientists try to mimic the in vivo niche by adding soluble FGF2.   When FGF2 is not present or withdrawn from cell culture strategies, stem cells now are able to “think” and start differentiating into other cell types. In some paradigms, researchers add fresh FGF2 every day, that means coming on Saturdays and Sundays to just maintain adequate FGF2 levels.

Recently, data from our lab showed that adding FGF2 every day might not be enough.  When we looked at FGF2 levels in stem cell culture, after 10 hours in culture, no more FGF2 was left.  When we graphed this over the course of one week, FGF2 levels looked like a roller coaster ride, starting high, going low, then spiking back up, before rapidly decaying again.  This result was extremely interesting, and very important because it proved that the current standard stem cell culture techniques using FGF2 were far from optimal.  In any given 24-hour period, where stem cells are without FGF2 for greater than half that time, they start to drift away from the stem cell state and begin the differentiation process.

To address this problem, our group came up with a way to keep FGF2 levels stable for at least 3 days in culture.  When we did this, we found that stem cells were maintained better in the stem cell state, and eliminated some of that “drift” and spontaneous differentiation.  This new product, StemBeads FGF2 allows for a savings of time, money savings from the use of fewer media changes, and most importantly, facilitates the growth of better cells.

So, while a lot of us enjoy the ups and downs of a roller coaster ride to get our adrenaline pumping, ups and down of FGF2 levels in standard stem cell culture techniques are far from optimal, and a paradigm shift may be in order.

If you enjoyed this post, please consider subscribing below to our monthly newsletter through which we provide stem cell scientists information on the most current research topics and tools to help them constantly improve their stem cell culture experience.
 

To Successful Research, 

StemCultures

Information for scientists by scientists on all things related to stem cell growth including stem cell culture, culture medium, tissue culture, fibroblast growth factor (fgf2) and more.

Note: StemCultures facilitates posting on this blog, but the views and accounts expressed herein are those of the author(s) or interviewee(s) and not the views or accounts of StemCultures its officers or directors whose views and accounts may or may not be similar or identical. StemCultures, its officers and directors do not express any opinion regarding any product or service by virtue of reference to such product or service in this blog.