Q&A with Dr. Aaron Gardner

[agardner]

dr. gardner,
you said the hair follicle including hair shaft in vitro of lauster's biochip reactor was tiny.
in 2009 they presented the tiny so-called microfollicle already. if it's still the case today, what happened during all these years? are you sure they didn't scale it up already?
we will hopefully see the presentation soon when desmond puts it online on the weekend.
will the presentation answer some questions and really represent the latest status, or did they present some old news to not reveal all their latest findings?

also, i was wondering, if you and jahoda would get a license maybe to use their biochip technology for your own research. i saw you stated that there is of course some sort of competition to keep the job and so on, but is there no chance to get in touch with team lauster and exchange some findings? they could profit of your and jahodas big knowledge about DP cells and culturing stuff, and in return you could proceed with your culturing experiments with an optimum environment from the biochip. i would see it more as a win-win situation than a hard competition between those two teams. because if the biochip is the only possibility to allow for optimum culturing, other teams would only get access to it through licensing because of the patents they hold.

but that's just an idea. i just can't see a way how the manual culturing in dishes with all these hanging drops techniques etc. can pan out to achieve fast access. if i understand it correctly, it's time consuming and has failure rates due to the fact that the dishes have to be monitored and supplied manually every few days.

i would love to see at least european labs working together like you/jahoda and TU berlin to bring this thing to the next level. there's also no risk for both teams to lose their job here =) because both teams are having invaluable know how.

I don't know how their research is progressing beyond what was presented at the conference. I'm not trying to put down their work when I call it a micro-follicle. It's amazing that they are able to generate these in a lab system, I would love to see how they develop after implantation.

But, I also don't think it's a good idea for groups to be replicating each others work, I think it's better if there are several groups trying several different techniques.

The manual culturing that I do is small scale, as this is all that is required for research purposes i.e I often only need 20 spheres for an experiment. There are technologies in place to automate and scale up the hanging drop system to produce the thousands of spheres that would be required for clinical use.

[walrus]

I’ve mentioned previously implanting these constructs into skin which has underlying problems will not “cure” baldness. Co-therapies limiting the degradation of any new follicles would be required.

Even if the DP cells are derived and multiplied from an apparently 'resistant' area of the scalp, i.e. the back of the head?

[agardner]

sdsurfin please explain the difference between inductivity retention and gene expression. I'm under the impression that they are essentially the same. I'm thinking that cellular hair "inductivity" refers to genetic instructions in the cell for those cells to morph into hair and "genetic expression" is merely the expression of those genes. These look to me like very similar concepts. It seems to me that the measurement of one would EQUAL the measurement of the other. In other words, if they have achieved 40% genetic expression then that also means they have achieved 40% hair inductivity. And if this is correct then that is an improvement over Dr. Jahoda's numbers late last year because back then he achieved 22% I think. So it looks to me like it's nearly doubled.

Can someone please explain to me why I'm right or wrong?

Several thousand genes are expressed differently in 2D DP cultures compared to a healthy functional DP. This change in gene expression results in the loss of inductive capacity of human DP cells after 2D culture.

By culturing the DP in 3D, after an initial 2D culture, 40% of the expression of those genes is restored to levels seen in a healthy functional DP.

This restoration of 40% of DP character partially restores the inductive capacity of the DP cells, 5 of 7 donors with a success rate of 10-60% (i.e 10 spheres implanted, 1 follicle develops = 10% success rate)

We believe that further restoration of the DP character will further increase the inductive capacity of culture DP cells.

[agardner]

from what we have read, Dr. Xu is going the other way over iPS cells to multiply the iPS cells in mass and then convert them rapidly into DP cells which should create full inductivity and full gene expression of the cells. what is your opinion about that way? is it 100% sure that if someone finds a solution for converting iPS cells to DP cells that these DP cells will really have the full inductivity and full gene expression? if not, dr. Xu is wasting his time here, and we all should warn him =D

See this figure for a nice explanation of the differences in technique:

http://www.nature.com/nature/journal/v460/n7251/fig_tab/nature08180_F2.html

iPS technology is attempting to de-differentiate any cell into a pluripotent (capable of producing any adult tissue) stem cell, and then re-differentiating these cells into a dermal papilla cell. Hopefully an inductive one.

We are attempting to restore the inductive capacity of DP cells that is lost, and alternatively try to differentiate DP like cells (e.g. dermal sheath cells) into inductive DP.

Both methods are equally valid ways of investigating what it is that makes an inductive DP cell, and any findings from one are applicable to the other.

[agardner]

about jahoda's experiment:

please everyone correct me if i'm wrong here:
we heard that jahoda combined epithelial and DP cells and implanted them into his arm (or was it his wife's arm?). and the cells developed into a normal hair then. i think arashi has some good info on that. arashi, maybe you can elaborate on that a bit. since we heard that story, we we're assuming that epithelial cells and dp cells are the only cells which are required for forming a hair follicle.
the important fact here: jahoda used fresh isolated cells which didn't go through multiplication (because they would lose their properties). so this was a proof of concept that those 2 cell types lead to the formation of a new follicle, IF the cells didn't lose their properties.

is that story right so far?
if so, is this experiment reproducable anytime? or what details are we missing here?
because if this were true the only puzzle to solve would indeed be to crack the code how to expand/multiply DP cells without losing their properties. and THIS is what we all thought was figured out by some teams all around the world and also presented on the hair congress.

and also important here: is THAT hair still growing on the mentioned arm? we would like to see that =)

This is the paper in question:

http://www.nature.com/nature/journal/v402/n6757/fig_tab/402033a0_F1.html

The cells were not expanded in culture. Expansion in culture is required for the large numbers of follicles that will be used clinically. This study was meant to demonstrate that intra-patient transplantation was possible without generating an immune response and that fresh dermal sheath cells were capable of inducing follicle.

[walrus]

Both methods are equally valid ways of investigating what it is that makes an inductive DP cell

Are there specific genes that you think may be crucial for inductivity (already known, or in theory)? I imagine they could be knocked down (using something like RNAi) in non-proliferated DP cells to examine their relative importance?

[agardner]

what? jahoda's DSC cells only? and in his wife's arm? what about rejection? why did jahoda do that? why not in his own arm?

and if DSC cells could do that trick alone, why not concentrate on multiplying those cells and inject them instead?

man, i'm totally confused now. i'm lost. it feels like the cure is here already but nobody is willing to go the final step =D

Because the study used fresh DS sections from inductive follicles. The cells weren't expanded in culture. This expansion step is when inductivity is lost, but is required to generate the large numbers of cells required to make thousands of follicles.

[Arashi]

Wow thanks for all the answers again Aaron !!

Because the study used fresh DS sections from
inductive follicles. The cells weren't expanded in culture. This expansion step is when inductivity is lost, but is required to generate the large numbers of cells required to make thousands of follicles.

But I think the question is then: is it equally hard to culture DSC cells als DP cells (while retaining enough genes exressed) ? Because from Jahoda's experiment it seems that Dermal Sheath cells are as good at forming new HF's as DP cells, right ?

[hellouser]

Because from Jahoda's experiment it seems that Dermal Sheath cells are as good at forming new HF's as DP cells, right ?

I certainly hope so, because if thats the case, than Replicel will have a serious shot at a solid treatment for baldness.

Aaron, any thoughts on Replicel's approach?

http://www.replicel.com/

[joachim]

Because the study used fresh DS sections from inductive follicles. The cells weren't expanded in culture. This expansion step is when inductivity is lost, but is required to generate the large numbers of cells required to make thousands of follicles.

i understand. expansion always leads to lost of the inductive properties.
if Replicel is working with those cells alone this is probably their core competence. so i assume they are already able to expand them. and maybe Dr. Xu could tell us if he also tried to create DSC cells via conversion of iPS cells.
because Dr. Xu is able to convert epithelial cells from iPS cells already and his next steps will be the other required cells. maybe it turns out, that DSC cells are easier to convert from iPS.

the important fact here is: DSC cells alone are also able to induce new hair follicle formation.

so in theory: if we would have an unlimited supply of inductive DSC cells only, and inject them into the scalp, then they are able to induce new hair follicles, with more or less efficacy. even if only 1 out of 10 injections (10% efficacy) would lead to new hair follicles, this treatment would be huge and practically a cure, considering that we could do the treatment multiple times. so this is what replicel is doing all the time, and if i remember correctly, they achieved 15 % average efficacy already in some of their studies. the questions about seb. glands and growing angle still remain. however, replicel seems more and more promising if we combine all these facts.

[hellouser]

i understand. expansion always leads to lost of the inductive properties.
if Replicel is working with those cells alone this is probably their core competence. so i assume they are already able to expand them. and maybe Dr. Xu could tell us if he also tried to create DSC cells via conversion of iPS cells.
because Dr. Xu is able to convert epithelial cells from iPS cells already and his next steps will be the other required cells. maybe it turns out, that DSC cells are easier to convert from iPS.

the important fact here is: DSC cells alone are also able to induce new hair follicle formation.

so in theory: if we would have an unlimited supply of inductive DSC cells only, and inject them into the scalp, then they are able to induce new hair follicles, with more or less efficacy. even if only 1 out of 10 injections (10% efficacy) would lead to new hair follicles, this treatment would be huge and practically a cure, considering that we could do the treatment multiple times. so this is what replicel is doing all the time, and if i remember correctly, they achieved 15 % average efficacy already in some of their studies. the questions about seb. glands and growing angle still remain. however, replicel seems more and more promising if we combine all these facts.

Requiring patients for repeat treatments would be costly. I can't imagine that one visit for a Replicel treatment will be cheap enough for Norwood 7s to do so many times to go back down to Norwood 1. The treatment SHOULD work once and right off the bat. However, required multiple sessions would ensure a constant cash flow; sort of like a subscription policy. One treatment should not cost as much as a hair transplant unless the amount given from a single session is equal to or more than a traditional transplant.

Notice though in Replicel's video here;

https://www.youtube.com/watch?v=cCe5mg7X6zg

They say that the DSC cells can rejuvenate existing follicles as well as induce NEW follicles.

This sounds pretty promising for lower norwoods.

[joachim]

Requiring patients for repeat treatments would be costly. I can't imagine that one visit for a Replicel treatment will be cheap enough for Norwood 7s to do so many times to go back down to Norwood 1. The treatment SHOULD work once and right off the bat. However, required multiple sessions would ensure a constant cash flow; sort of like a subscription policy. One treatment should not cost as much as a hair transplant unless the amount given from a single session is equal to or more than a traditional transplant.

Notice though in Replicel's video here;

https://www.youtube.com/watch?v=cCe5mg7X6zg

They say that the DSC cells can rejuvenate existing follicles as well as induce NEW follicles.

This sounds pretty promising for lower norwoods.

their demo video is pretty cool. and yes, rejuvenating and also creating NEW hairs.
and phase 2 will start this year in Germany it says on their website. dosing is important. and believe me, if there is a cure, nobody would care about going in for multiple treatments. the costs would come done quickly. they are also developing special instruments to inject the cells. i assume you can make 1000 injections per hour easily. the mass culturing of cells will be probably automated somehow. they say it takes 3 months for cell culturing.
however, a subscription based model would also be ok. i wouldn't mind spending some thousand dollars a year IF it's a proven method which keeps existing hair AND gives 20% more density with each treatment.

i'm getting more and more faith in Replicel now. there's a backed science behind it.

interesting: rolf hoffmann is a German, and founded replicel with a colleague in 2003.

[micgeed]

Mr. Gardner I have a few important ques. #1 Will hair cloning work for someone with a Norwood 6 or 7? And if so, will it restore a full head of head, with a normal hairline as well. #2 If all that worked out, once you are finished with the procedure, you won't have to go back ever again? #3 In the future when you develop gray hair will the cloned hair turn gray as well? #4 Last but not least how much do you think this would cost for a person with a Norwood 6 or 7, BALLPARK/EDUCATED GUESS?? Thank you, Mr. Gardner for you time.

[joachim]

i just noticed something: working with DSC cells by just injecting them could truely be effective because: compared to the dermal papilla which is surrounded by all other cells the dermal sheath cup is accessible from outside. thus, it's relatively easy for the injected DSC cells to attach to the existing dermal sheath cup. but if you inject DP cells they are not able to find their way to existing dermal papilla because they can't go through the other cell walls like DSC and outer root sheath. this is probably the reason why nigam's and other experiments failed when they tried to rejuvenate the follicles with the use of 2D cultured dp cells.
does that makes sense? your opinions, please.

seems like DSC is the new key to success.
Replicel in combo with Histogen could be awesome!
replicel phase 2 results are important now.

[nameless]

At the moment they would come out un-pigmented. Incorporation of patient melanocytes (the cells that provide the colouration) is something that is in the works.

Couldn't we just dye the new hair?

I think that if you guys can just figure out how to get us the hair we will just dye it to get color.