Final Days: Chinese Scientists Have Solved the DP Culturing Problem! (2014)

[clarence]

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[534623]

Japanese Scientists (Tsuji Lab) vs. Chinese Scientists

So I'm pretty sure he'll be negative about this too (since it's not HASCI, lol)

That means you can find something positive within all in this thread mentioned trash science - sorry, but I cannot:

Scientists from the Tsji-Lab
The pic below shows what scientists from Tsuji's Lab had as source for the REARRANGEMENT of stem cells and their niches, to produce at least 1 fully functional human hair-bearing follicle after intracutaneous transplantation into the back skin of a nude mouse ...


... to get at least this:

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"Bioengineering of human hair follicle germ
This procedure was performed with the approval of the ethics board of the Tokyo University of Science and Tokyo Memorial Clinic. Small pieces (1.0–2.0 cm2) of occipital scalp skin were obtained from 39- and 63-year-old male donors, who provided informed consent in accordance with the Helsinki Declaration. The scalp samples were treated with povidone-iodine and 70% ethanol for 10 s, twice successively.

- The hair follicles in the anagen phase were then dissected at the upper end of the hair bulb, using a surgical knife.

- The intact DPs and bulge-derived epithelial cells were isolated as described above.
- The bioengineered hair follicles were then reconstituted using human intact DPs and the bulge region-derived epithelial cells, by using the organ germ method7.
At 21 days after transplantation with the inter-epithelial tissue-connecting plastic device, the growth of pigmented hair shafts was observed underneath the epithelium of the host skin. The hair shafts were recovered by cutting into the host epithelium."
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So 1 ingrown hair (see their pic above or within their paper), but at least 1 fully functional hair-bearing follicle.

Chinese scientists

The pic below shows what the Chinese scientists had as source ...


... to produce this:

[...]
We are the first to show that passage 3 human hair germinal matrix cells differentiate into hair-like fiber in the presence of human DP spheroids.

These results suggest that three-dimensional Matrigel culture technique is an ideal culture model for forming DP spheroids and that sphere formation partially models the intact DP, resulting in hair induction, even by high passage DP cells.

http://online.liebertpub.com/doi/abs....TEA.2013.0547

... a "hair-like fiber".

So the question remains why the scientists from the Tsuji-Lab could, at least, produce 1 normal fully functional hair-bearing follicle and the Chinese scientists just a "hair-like fiber"?
It has something to do with the pics with circles and numbers I posted above; the source the Tsuji-Lab scientists used contained, at least partially, the proper instructions for the proper "rearrangement" of stem cells and their niches - what finally resulted in a fully functional hair follicle. Sorry, but with the given and sufficient "data" (=genetics) you can see in the 1st pic, YOU know also which numbers should be there in the empty circles b/d/f and h. - and the source they used "knew" this too.

The Chinese scientists "failed" insofar (it wasn't their intention to produce with this approach fully functional hair follicles) and produced just "hair-like fibers", because their souce lacked the proper instructions (what the hell should I build/create?) - see circles pic #2: With the given instructions, as you can see them in the 2nd pic, the souce could produce either nothing or something else.

In simple words, the more you add "proper instructions" into the mix, the better a given souce knows what exactly it should create - even if they got just parts of instructions. To replicate/multiply THE "proper instructions" is still THE challenge for scientists; today, it's easy to replicate/multiply cells, but very difficult to replicate -let's say - the "pilot", who instructs all the different cells. The Tsuji-Lab scientists added at at least essential and natural "pilots" into the mix; but that means also that no "hair multiplication" took place, because they didn't and couldn't multiply the "pilots" - they just added them from a certain (limited) source.

[Tenma]

Thanks, IM. Really great work man

Didnt know until now the big "genetic gap" between adult dp cells used by tsuji lab and the ones cultured by the chinese investigators.

So, the chinese study is just about the good replication capabilities of Matrigel rather than the "quality" of the DP cells obtained

In a nutshell, replication of DP cells isnt the biggest issue right know. Its almost all about retaining the genetic instructions of the original cells.

[534623]

In a nutshell, replication of DP cells isnt the biggest issue right know. Its almost all about retaining the genetic instructions of the original cells.

Basically - correct. More specific "retaining the genetic instructions of the cells' NICHE - namely, the cells' natural environment.

The Chinese scientists mentioned in their paper that they used hair germinal matrix cells and the DP spheres; simply to examine the hair-inducing ability of their by them seperate produced DP spheres.

Hair germinal matrix cells?
I used the following pic from this website ...

http://www.sciencellonline.com/site/productInformation.php?keyword=2410

... for further explanations:

*FULL SIZE* http://www.sciencellonline.com/site/...man/2410_3.jpg

Besides NOT using the proper material for the production of fully functional hair follicles (scientists from the Tsuji-Lab used several COMPLETE/INTACT dermal papillae -meaning including the proper "pilot"- plus real hair follicles (stem)cells derived from the bulge area of hair follicles), as already mentioned, the Chinese scientists just used artificial DPs (meaning, the genetically proper "pilot" is missing - what is not so by using natural INTACT DPs) plus hair germinal matrix cells - also without their genetically proper "pilot"; which you can see in the pic as the green stuff (fibronectin = genetically proper "pilot" who instruct, in this case, matrix cells (= blue dots in the pic above).

So again, you can replicate/multiply cells (for example the blue labeled dots in the pic above) as much as you want and you can inject them into (bald) skin as much as you want - you will not get what you want without the extremely difficult to reproduce green stuff: the (stem)cells natural "niche". THIS stuff instructs cells what to do, how often they should divide, what the should build/create in general, etc etc etc. Stuff like the ACell or Matrigel, such stuff contains also instructions - but not the proper instructions; simply because they do not contain the proper DNA; their material is derived from something else; either produced on a synthetic basis or are derived e.g. from pig bladder ECM or something else). Such stuff simply CAN'T contain "proper instructions" - to simplify the terminology for this issue.

[Arashi]

You misunderstood some things Ironman. Tsuji lab wanted to show that it is possible to bio engineer human hair grafts producing human hair. They just took existing cells and hair grew. Those Chinese wanted to prove it's possible to culture dp cells without losing gene expression. They were the first in the world to do that. The next step would be to repeat Tsuji's experiment but this time with cultured cells.

[Arashi]

Also you overcomplicate things. What tsuji did is take 2 types of stem cells. Epithelial and mesenchymal. Those last were extracted from human dp. Mesenchymal = dp cells (or the other way around actually. Dp = subtype mesenchymal). That's the only part they used from the human dp.

So again the next step is to repeat what tsuji did only this time not with dp cells extracted from human dp, but with cultured ones

[Tenma]

without losing gene expression.

According to IM analysis they couldnt accomplish this part, Arashi.

[Arashi]

According to IM analysis they couldnt accomplish this part, Arashi.

Well that's what they did and that's why it's so big. Ironman seems to misunderstood several things.

[534623]

You misunderstood some things Ironman.

Fortunately, you are the one understands everything correct ...

Yeah they grew cosmetically correct HUMAN hair on mice, as you can see on the photo's.

Pffffft ...

You misunderstood some things Ironman. Tsuji lab wanted to show that it is possible to bio engineer human hair grafts producing human hair.

Did I say something different?? Oops, I forgot:
They tried to bio-engineer a penis on a mouse tooth.

They just took existing cells and hair grew.

Really? Scientists from Tsuji's lab? Just existing human cells?
Come on, learn reading ...

Those Chinese wanted to prove it's possible to culture dp cells without losing gene expression.

Did I say something different?? Not really ... learn also reading posts.

They were the first in the world to do that.

Perhaps for a newbie like you - but not for me or other scientists in this field.

So please, don't try to tell ME what they tried to accomplish and what material they used for their different experiments. AND, if you really find something wrong within my post, next time QUOTE these parts and PROVE me wrong with hard facts.

[Arashi]

Btw tsuji extracted the dp from the follicle itself, I said otherwise earlier. But fact remains all that about drivers and stuff Ironman talks about is irrelevant. An exact copy of dp cells should be able to achieve exactly the same as the existing counterpart

[Arashi]

Haha you are funny Ironman. I am not at home right now but will prove everything with source references when I am at home. You totally misunderstood everything like we are used from you

[534623]

Btw tsuji extracted the dp from the follicle itself, I said otherwise earlier.

Let me look ...

You misunderstood some things Ironman. Tsuji lab wanted to show that it is possible to bio engineer human hair grafts producing human hair. They just took existing cells and hair grew.

So according to your IQ and all my misunderstandings, every lab out there can simply took existing cells - and hair grows. That's nice. Fantastic. I'm really pissed that I never had this simple idea and protocol. Damn ...

Arashi, do you really think that all the other guys on these forum suffer also from ADHD like you?

Personally, I think you should put yourself into the "just reading" mode. Would be more helpful not just for you only - for many others too. Just a good advice ...

[Arashi]

I am going to use this document as my reference: http://www.tsuji-lab.com/en/pdf/Toyo...ncomms1784.pdf

Fortunately, you are the one understands everything correct ...Pffffft ...

Not sure what you mean but do those hairs to you look like normal mouse hair ? Tsuji did 3 their hair bio engineering experiment for 3 types of hair:
1) Mouse pelage hair
2) Mouse vibrissa hair
3) Human hair.

About that last: "Furthermore, the human bioengineered hair follicle germ, which was composed of the dissociated bulge region-derived epithelial cells and scalp hair follicle-derived intact DPs of an androgenetic alopecia
patient, grew a pigmented hair shaft in the transplantation area within 21 days after intracutaneous transplantation into the back skin of nude mice (Fig. 2b)"

AND

"By analysing the nuclear morphology with Hoechst staining40, we confirmed that the cells in
the bioengineered hair follicle were of human origin (Fig. 2b) "

Really? Scientists from Tsuji's lab? Just existing human cells?
Come on, learn reading ...

It's obviously you who can't read. Again: "Furthermore, the human bioengineered hair follicle germ, which was composed of the dissociated bulge region-derived epithelial cells and scalp hair follicle-derived intact DPs of an androgenetic alopecia patient , grew a pigmented hair shaft in the transplantation area within
21 days after intracutaneous transplantation into the back skin of nude
mice

Please re-read Tsuji's paper you didnt understand 1 single thing it seems. Tsuji combined existing human DP cells + epethelial cells and grew hair that way. That's all there is to it ! Now those Chinese have proven that they can culture DP cells. They need to repeat what Tsuji did now, as the next step.

[Arashi]

So the question remains why the scientists from the Tsuji-Lab could, at least, produce 1 normal fully functional hair-bearing follicle and the Chinese scientists just a "hair-like fiber"?

This pretty much sums it up. You just didn't understand it at all. Again, Tsuji lab wanted to demonstrate how to grow human hair out of some exisiting cells and they succeeded. Those Chinese wanted to show they could culture DP cells while preserving gene expression (so succeeding where Jahoda failed) ! Totally different objective. To show the DP cells didnt lose their follicle inducing ability (like normally happens when culturing DP cells), they even induced some hair like fibers. But all in a dish !! This is TOTALLY different than growing REAL hair (like Tsuji did) on skin. This doesnt somehow show the Chinese failed (like you make it look) at all. It just showed that the DP cells retained their HF inducing capability after culturing them. Now, again, the next step would be to repeat what Tsuji did but this time using CULTURED dp cells, instead of using existing ones taken from human donors. But if the DP cells really are the same as their original counterpart (and not have lost gene expression), then there's theoretically no reason why that wouldnt work.

[Arashi]

(or better: they now need to repeat Jahoda's last year's experiment and try it on human skin ! Jahoda grew hair but 78% gene expression was lost and the hair didnt have the right cosmetic features as a result. Theoretically that should now be solved !)