Hair Cloning with Autologous Cells - Bernstein Medical - Center for Hair Restoration
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New research published in the journal Developmental Cell has confirmed the importance of dermal sheath stem cells in maintaining the hair growth cycle. ((Rahmani W, et al. Hair Follicle Dermal Stem Cells Regenerate the Dermal Sheath, Repopulate the Dermal Papilla, and Modulate Hair Type. Dev Cell. 2014 Dec 8;31(5):543-58.)) These cells, located around the lower portion of growing follicles, form the basis of an experimental treatment, being developed by Replicel Life Sciences, Inc., to regenerate hair-producing follicles. If successful, the treatment will be a game-changer for the hair restoration industry.

Colony of self-renewing dermal sheath cellsColony of self-renewing dermal sheath cells

The study, published in December 2014, sought to confirm what had been indirect evidence of a type of stem cell residing in the dermal sheath (DS) that was said to replenish dermal papilla (DP) cells. The authors of the study suggest that they now have definitive evidence that new DP cells are derived from stem cells in the dermal sheath “cup” (DSC). This development clarifies the relationship between the DS and the DP and confirms that DSC cells play a critical role in hair follicle regeneration by repopulating the dermal papilla cells at the end of the telogen (resting) phase of the normal hair cycle.

Importance of the Dermal Sheath Cup Cells

The number of dermal papilla (DP) cells in a hair follicle has been found to be a determining factor as to when the anagen (growth) phase of the hair cycle is initiated. ((Chi W, Wu E, Morgan BA, et al. (2013). Dermal papilla cell number specifies hair size, shape and cycling and its reduction causes follicular decline. Development 140, 1676–1683.)) The gradual loss of DP cells over time results in a longer delay in the onset of the anagen phase; a longer telogen (resting) phase; and a hair follicle that shrivels and eventually disappears. This process, called miniaturization, plays out over multiple hair cycles and has been shown to be the primary contributor to androgenetic alopecia and eventual baldness. ((Randall VA. (2008). Androgens and hair growth. Dermatol. Ther. 21, 314–328.))

While dermal sheath cup (DSC) stem cells are known to be long-lived and self-renewing, it is not fully understood how they replicate or why the pool of DSC cells becomes depleted over time. We do know, however, that the gradual loss of DSC cells results in a failure to produce the necessary number of DP cells. And without enough DP cells to trigger the anagen phase, the follicle begins to miniaturize. It is clear that maintaining the population of DSC cells after each iteration of the hair cycle is very important in preserving and maintaining healthy and mature terminal hairs.

Replicel Reacts to the Study

The new data confirming the importance of the dermal sheath cup (DSC) cells was celebrated by researchers and executives at Replicel Life Sciences, Inc., who have been studying this issue for over a decade. Replicel is set to start phase II clinical trials of RCH-01, their proprietary treatment for androgenetic alopecia.

In the RCH-01 treatment, cloned DSC cells are injected into balding areas of the scalp where they are expected to reverse miniaturization and regenerate healthy, hair-producing follicles. Phase I trials of RCH-01, the results of which were published in 2012, showed that the treatment could produce promising results and that it was safe to administer. Six months after patients were treated with RCH-01, overall hair density increased by an average of 11.8% in ten patients out of 16. In two patients, overall hair density increased by more than 19%. There were no significant adverse safety events recorded. ((Lortkipanidze, N. Safety and Efficacy Study of Human Autologous Hair Follicle Cells to Treat Androgenetic Alopecia. In Clinicaltrials.gov. Retrieved July 26, 2012.)) Phase II clinical trials are set to begin in 2015, with data collection continuing for 39 months.

Through a 2013 agreement with Replicel, Japanese cosmetics giant Shiseido may introduce RCH-01 into the Asian market as early as 2018.

Image c/o Developmental Cell 31, 543–558, December 8, 2014 ª2014 Elsevier Inc.

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RepliCel Life Sciences; a company out of Vancouver, Canada; is studying the use of hair cloning techniques to treat male pattern baldness and hair loss in women.

The study is in progress, but analysis of the 6-month interim results of the first phases has been published. The preliminary results at 6 months show that almost two-thirds of subjects (10 out of 16, or 63%) received a greater than 5% increase in hair density at the injection site. Of that group of 10 subjects, seven of them saw hair density improve by more than 10%. In one subject vellus hair density increased 24.9%, terminal hair density increased 14.5%, overall hair density increased by 19.2%, and cumulative thickness per area increased by 15.4%. There were no significant adverse safety events reported in the first 6 months of the trial.

Phase I/IIa of the RepliCel study involved injecting male and female subjects with their own (autologous) dermal sheath cup cells (DSCC), which were replicated or cloned using RepliCel’s laboratory technology. A preliminary analysis of the safety of the injections, as well as a preliminary analysis of the efficacy of the treatment in growing hair, was announced in May 2012 and presented to the European Hair Research Society in June 2012. Subjects in this part of the study will continue to be monitored for any adverse physical reactions and to assess hair growth at 12 months and 24 months after treatment.

Phase IIb of the study is designed to help the RepliCel researchers formulate the optimal treatment for hair growth. Some of the treatment regimens that will be tested include the use of different concentrations of cells and different treatment schedules, plus the effects of single injections versus repeat injections. The final protocols for Phase IIb are currently being worked out, with the clinical trial expected to begin in late 2012.

Reference:

Lortkipanidze, N. Safety and Efficacy Study of Human Autologous Hair Follicle Cells to Treat Androgenetic Alopecia. In Clinicaltrials.gov. Retrieved July 26, 2012, from http://clinicaltrials.gov/ct2/show/NCT01286649.

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RepliCel Life Sciences; a company based in Vancouver, Canada; is investigating hair cloning techniques in order to develop a treatment for androgenetic alopecia, or common genetic hair loss.

Research conducted by the company’s scientific founders and lead scientists, Drs. Kevin McElwee and Rolf Hoffmann, has shown that a certain type of cell, called a dermal sheath cup cell, is integral in initiating the growth of mature hair follicles. ((McElwee KJ, Kissling S, Wenzel E, Huth A, Hoffmann R (2003) Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol 121: 1267–1275.)) This mechanism of follicle growth, when coupled with previous research on dermal papillae cells, is key to our understanding of hair loss and is a potential avenue for developing a treatment that could reverse hair loss.

In their 2003 study, “Cultured Peribulbar Dermal Sheath Cells Can Induce Hair Follicle Development and Contribute to the Dermal Sheath and Dermal Papilla,” the scientists found that the dermal sheath cup cells are the “reservoir” of stem cells that control both the hair growth cycle of a follicle and formation of new hair follicles.

These breakthrough findings led to RepliCel’s seeking patents for their proprietary process of isolating and preparing dermal sheath cup cells for the treatment of hair loss. Patents have been issued in Europe and Australia, and are currently pending in the US, Canada, and Japan.

In 2012, RepliCel is studying the safety and efficacy of hair regeneration from autologous dermal sheath cup cells. In the study, cells will be harvested from patients, replicated in a laboratory, and then injected into a balding area to determine if the treatment will stimulate the growth of new hair follicles in what was a bald area.

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Scientists from Durham University in the UK have shown for the first time that a lab technique, called a three-dimensional cell culture, can produce spherical structures that are similar to naturally occurring structures in hair follicle formation (called dermal papilla or DP). This breakthrough study by Claire Higgins and Colin Jahoda, published in the June 2010 issue of the journal Experimental Dermatology, ((Higgins C, Jahoda C, et al. Modelling the hair follicle dermal papilla using spheroid cell cultures. Experimental Dermatology 2010; 19: 546–548.)) has the potential to unlock the ability of researchers to develop functional DP cells which can be used in hair restoration techniques such as hair cloning or hair multiplication.

Background

Hair cloning techniques have been theorized for decades. The basic idea is:

  1. a physician takes a sample of skin cells from a patient
  2. dermal papilla cells are extracted
  3. the DP cells are cloned (multiplied) in a laboratory culture (i.e., a petri dish)
  4. the cell formation is then injected back into the patient’s balding scalp where it produces permanent hair that continues to grow

The first three steps are a piece of cake. But that is when the strategy breaks down. When DP cells are grown in a petri dish they exhibit some of the qualities of DP cells in the human body but not all, so injecting this aggregate into the skin fails to lead to hair follicle growth. Something was missing.

In 1991, Wobus, et al published a study in the journal Differentiation ((Wobus AM, Wallukat G, Hescheler J. Differentiation 1991: 48: 173–182.)) that described a new technique for researching cells that in nature exist as clumps or masses of cells. The idea was to suspend a group of cells under a flat surface so that gravity would pull the cells into a droplet. This “hanging drop” method yielded a three-dimensional culture that enabled the study of embryonic stem cells as well as the proteins they produce that allow for intercellular communication.

Having hit the wall with two-dimensional DP cultures, Higgins and Jahoda set out to try Wobus’ concept of using 3-D cultures to study DP cells.

The Study

Higgins and Jahoda harvested eight cell strains of human DP cells taken from scalp hair follicles. These eight strains were cultured in either 35-mm dishes or hanging drop cultures consisting of 3,000 cells each. The cultures were maintained between 30 and 72 hours, then collected and analyzed using immunofluorescence or transcriptional techniques.

Results

The DP cells grown in hanging drop, 3-D cultures exhibited behavior significantly akin to DP in human hair follicles. The 2-D cultures grown in the 35-mm dishes did not.

Conclusion

Without the ability to form functional dermal papilla aggregations, hair cloning was essentially at a dead end. In the 3-D configuration, the aggregated cells were able to communicate with one another and to continue to differentiate as hair follicles. By using Wobus’ 3-D hanging drop technique, Higgins and Jahoda may have unlocked the secret to forming these powerful, but elusive, structures that are critical to the hair growth cycle.

Following this study, more research needs to be performed to induce the spherical cells to initiate the growth of new hair follicles and to develop ways to ensure that the induced hair follicles are immune from the factors that cause genetic hair loss. Should those two riddles be solved, hair loss will have been effectively cured.

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