Higgins Lab - Methods For The Isolation And 3d Culture Of Dermal Papilla Cells From Human Hair Ap

[Noisette]

Hi Guys,
It has been a while since my last post.
I've seen this study (from Higgins Lab) on Research Gate, I wonder if you have read it ? I upload the PDF

Higgins Lab : http://www.higginslab.org/people

Title : Methods for the isolation and 3D culture of dermal papilla cells from human hair

Abstract: " The dermal papilla is a cluster of mesenchymal cells located at the base of the hair follicle which have a number of important roles in the regulation of hair growth. As a consequence, in vitro models of these cells are widely used to study the molecular mechanisms which underlie hair follicle induction, growth and maintenance. While dermal papilla from rodent hair follicles can be digested prior to cell isolation, the unique extracellular matrix composition found in human dermal papilla renders enzymes such as trypsin and collagenase insufficient for digestion of the dermal papilla into a single cell suspension. As such, to grow human dermal papilla cells in vitro, the papilla has to first be isolated via a micro-dissection approach from the follicle. In this article we describe the micro-dissection and culture methods, which we use within our laboratory, for the study of human dermal papilla cells ".

To summarise: here we have detailed methods on human dermal papilla isolation, and 3D spheroid culture. The dermal papilla is an indispensable component of the hair follicle that plays a crucial inductive role in hair growth. Isolation and culture of these cells can be arduous and time-consuming, and dramatic transcriptional shifts occur in culture, decreasing our ability to gain insight to the role of the papilla in hair growth and cycling with in vitro models. Culture of papilla cells in 3D spheroid structures in vitro may provide a useful tool for hair research going forward, and hopefully aid in the discovery of new therapeutic targets for hair growth.

Some screenshots:

 

[MrV88]

I wonder when they will just say "research done come and get your treatment, make us billionaires you bald morons"

 

[PrinceWilliamThe2nd]

@ Noisette: Wasn't the culturing of papilla cells already solved by Tsuji and coworkers? I think the major issue is still the cultivation of epithelial stem cells, right?

 

[lemoncloak]

Is it just me or is everyone successfully growing DPs these days? I ain't complaining.
It's always nice to log in and see posts like this.
One more thing, they say "While it is known that hair follicle stem cells in miniaturising follicles have a reduced ability to convert into stem cell progenitors, it is not known whether this is due to a lack of signalling from the dermal papilla, or an inability of these cells to respond to signals from the dermal papilla." Basically we don't know if the epithelial stem cells are harmed by Androgenetic Alopecia which would explain why everyone (except Tsuji) is concentrating their efforts on the lowest common denominator, the DP (tho personally I think it's safer to put the two types of cells together to form structures before injecting, rather than hope they will communicate and form in vivo as they -sort off- naturally do)

 

[Kev123]

I won't pretend like I know what I just read so I will ask: How does growing DP cells alone, help stop male pattern baldness?

Stop me if i'm wrong, but from what I understand from this wikipedia explanation of what DP does "Blood vessels in the dermal papillae nourish all hair follicles and bring nutrients and oxygen to the lower layers of epidermal cells." this is needed to keep scalp and follicles healthy, but does it stop your hair follicles from dying due to DHT?

Or is this study not even supposed to stop hair loss but instead just help in finding out ways to regrow hair?

Thanks, I suck at this "advanced" biology stuff. (It's advanced to me).

 

[cocona]

I won't pretend like I know what I just read so I will ask: How does growing DP cells alone, help stop male pattern baldness?

Stop me if i'm wrong, but from what I understand from this wikipedia explanation of what DP does "Blood vessels in the dermal papillae nourish all hair follicles and bring nutrients and oxygen to the lower layers of epidermal cells." this is needed to keep scalp and follicles healthy, but does it stop your hair follicles from dying due to DHT?

Or is this study not even supposed to stop hair loss but instead just help in finding out ways to regrow hair?

Thanks, I suck at this "advanced" biology stuff. (It's advanced to me).

So the entire paper is about the protocol their lab follows for isolating and culturing dp(dermal papilla) cells.

So basically you have a bunch of epithelial stem cells inside a shroud called the dermal papilla which is made of mesenchymal stem cells. The dermal papilla signals the epithelial stem cells to create new hair. In androgenic alopecia once a hair miniaturizes sufficiently it will get stuck in the telogen(Resting) phase and no longer grow hair. It is currently unknown if this is because the dermal papilla loses the ability to signal new hair growth of if the stem cells inside the papilla lose their ability to receive that signal.

Their goal is to culture dp cells (mesenchymal stem cells). The problem is when done for human dp cells those cells lose the ability to signal the stem cells to grow new hair quickly. An interesting part is they use a spherically shaped culture which allows them to partially retain the dp cell's ability to signal new hair growth in the stem cells.

Sidenote1: Tsuji is also working on solving hte problem of culturing mesenchymal stem cells without losing their ability to induce new hair growth in epithelial stem cells.
Sidenote2: The working hypothesis is that if you culture dp cells and epithelial cells from the donor area then implant them on the top of the head then they will not miniaturize from dht.

 

[mr_robot]

This is quite interesting although it acknowledges they can only get partial restoration of inductivity.

What is more important about this paper is that methods of seperating the DP cells from the biopsy which is currently done in a very skilled labour intensive process can be automated, this should make any potential treatment significantly cheaper.

 

[Trichosan]

My conclusion is they have developed a good research tool to further investigate the unknown factors of hair growth. It brings more eyes on the problem. That's all. But that, in itself is positive.

 

[hairblues]

@Noisette what do you think of PRP and dermal papilla?

from a study https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4622412/

PRP has been reported to induce the proliferation of dermal papilla cells by upregulating fibroblast growth factor 7 (FGF-7) and β-catenin, as well as extracellular signal-related kinase (ERK) and Akt signaling [2]. Anagen-associated angiogenesis has been suggested as one of the important factors in active hair growth, because of the secretion of VEGF by the keratinocytes of the outer root sheath and fibroblasts of the dermal papilla [6, 7]. Increased secretion of VEGF influences growth of normal and pathological dermal structures [8]. Tobin et al. [9] reported the hair follicle mesenchyme exhibits significant hair cycle-associated plasticity. Modulation of these cell interchanges is likely to be important during clinically important hair follicle transformations (e.g., vellus to terminal and terminal to vellus during androgenetic alopecia [9]). Injection of PRP has been demonstrated to improve cutaneous ischemic conditions and to increase vascular structures around hair follicles [10]. Many of the current treatment modalities for pattern hair loss have been shown to modulate angiogenesis and enhance blood flow [11]. We aimed to clarify the effects of PRP scalp injection in humans affected by androgenic alopecia. It could improve hair regrowth, thereby making the PRP procedure an alternative to finasteride or minoxidil. The data we report demonstrate the clinical efficacy and histological safety of PRP treatment. Moreover, patients’ satisfaction and computerized trichogram analysis have confirmed the quality of the results.

thinking to try PRP with or without ACELL to mostly maintain and hopefully thicken some strands NOT as a miracle cure...

Do you have thoughts on it? I respect your opinion.

 

[Noisette]

@ Noisette: Wasn't the culturing of papilla cells already solved by Tsuji and coworkers? I think the major issue is still the cultivation of epithelial stem cells, right?

Yes You're right The Major issue (september 2016) for Tsuji's lab is to cultivate epithelial stem cells.

@Noisette what do you think of PRP and dermal papilla?

from a study https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4622412/

PRP has been reported to induce the proliferation of dermal papilla cells by upregulating fibroblast growth factor 7 (FGF-7) and β-catenin, as well as extracellular signal-related kinase (ERK) and Akt signaling [2]. Anagen-associated angiogenesis has been suggested as one of the important factors in active hair growth, because of the secretion of VEGF by the keratinocytes of the outer root sheath and fibroblasts of the dermal papilla [6, 7]. Increased secretion of VEGF influences growth of normal and pathological dermal structures [8]. Tobin et al. [9] reported the hair follicle mesenchyme exhibits significant hair cycle-associated plasticity. Modulation of these cell interchanges is likely to be important during clinically important hair follicle transformations (e.g., vellus to terminal and terminal to vellus during androgenetic alopecia [9]). Injection of PRP has been demonstrated to improve cutaneous ischemic conditions and to increase vascular structures around hair follicles [10]. Many of the current treatment modalities for pattern hair loss have been shown to modulate angiogenesis and enhance blood flow [11]. We aimed to clarify the effects of PRP scalp injection in humans affected by androgenic alopecia. It could improve hair regrowth, thereby making the PRP procedure an alternative to finasteride or minoxidil. The data we report demonstrate the clinical efficacy and histological safety of PRP treatment. Moreover, patients’ satisfaction and computerized trichogram analysis have confirmed the quality of the results.

thinking to try PRP with or without ACELL to mostly maintain and hopefully thicken some strands NOT as a miracle cure...

Do you have thoughts on it? I respect your opinion.

Hey Hairblues

I hesitate to give my point of view because, maybe, my thought on this treatment is misleading

I am skeptical about this treatment (for men) because, over the years, I don't see and read some positive testimonials about PRP on androgenetic alopecia (the balding areas). But, some guys have some results like an increase in hair density on thinning area.

It seems that for women, PRP + ACELL is a better treatment than PRP only.

PRP + ACELL from Doctor Cole is maybe a good treatment for women (thinning areas). On his study, women respond to this treatment with an increase on hair density

@Swoop : Hello Swoop, What do you think about this ? thanks !

 

[Mach]

@Noisette what do you think of PRP and dermal papilla?

from a study https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4622412/

PRP has been reported to induce the proliferation of dermal papilla cells by upregulating fibroblast growth factor 7 (FGF-7) and β-catenin, as well as extracellular signal-related kinase (ERK) and Akt signaling [2]. Anagen-associated angiogenesis has been suggested as one of the important factors in active hair growth, because of the secretion of VEGF by the keratinocytes of the outer root sheath and fibroblasts of the dermal papilla [6, 7]. Increased secretion of VEGF influences growth of normal and pathological dermal structures [8]. Tobin et al. [9] reported the hair follicle mesenchyme exhibits significant hair cycle-associated plasticity. Modulation of these cell interchanges is likely to be important during clinically important hair follicle transformations (e.g., vellus to terminal and terminal to vellus during androgenetic alopecia [9]). Injection of PRP has been demonstrated to improve cutaneous ischemic conditions and to increase vascular structures around hair follicles [10]. Many of the current treatment modalities for pattern hair loss have been shown to modulate angiogenesis and enhance blood flow [11]. We aimed to clarify the effects of PRP scalp injection in humans affected by androgenic alopecia. It could improve hair regrowth, thereby making the PRP procedure an alternative to finasteride or minoxidil. The data we report demonstrate the clinical efficacy and histological safety of PRP treatment. Moreover, patients’ satisfaction and computerized trichogram analysis have confirmed the quality of the results.

thinking to try PRP with or without ACELL to mostly maintain and hopefully thicken some strands NOT as a miracle cure...

Do you have thoughts on it? I respect your opinion.

If you got the money it makes perfect sense. If you don't then don't get it. I'd go to Greco or Cole

 

[hairblues]

Yes You're right The Major issue (september 2016) for Tsuji's lab is to cultivate epithelial stem cells.



Hey Hairblues

I hesitate to give my point of view because, maybe, my thought on this treatment is misleading

I am skeptical about this treatment (for men) because, over the years, I don't see and read some positive testimonials about PRP on androgenetic alopecia (the balding areas). But, some guys have some results like an increase in hair density on thinning area.

It seems that for women, PRP + ACELL is a better treatment than PRP only.

PRP + ACELL from Doctor Cole is maybe a good treatment for women (thinning areas). On his study, women respond to this treatment with an increase on hair density

@Swoop : Hello Swoop, What do you think about this ? thanks !

That was my thoughts to I have read more positive about women reviews then men...the men seem to be mostly disastrous. Women i have read positives for maintaining and getting some hair caliber improvement.
I read somewhere it has to do with inflation which most women have..I have some inflammation forgot the name of the cells when they did a biopsy but at first they misdiagnosed me with areata because of the inflammation cells...my DR was excited when I told him because he said the people with inflammation did better with PRP...
Of course it;s a big chunk of change to do when its still a crap shoot..he told me 2/3 do well 1/3 non responder.

 

[That Guy]