Cellular therapy with human autologous adipose-derived adult cells of stromal vascular fraction for alopecia areata
Anderi et al. Stem Cell Research & Therapy
Cellular therapy with human autologous adipose-derived adult cells of stromal vascular fraction for alopecia areata
Rami Anderi 1 2
Nehman Makdissy 0
Albert Azar 4
Francine Rizk 3
Aline Hamade 3
0 Department of Biology, Faculty of Sciences 3, Lebanese University , Kobbe , Lebanon
1 Stem Cells, Organogenesis and and Regenerative Medicine, Lebanese University , Beirut , Lebanon
2 Cosmetic Plastic Surgery Center , Beirut , Lebanon
3 Department of Biology, Laboratory of Therapeutic Innovation, Faculty of Sciences 2, Lebanese University , Fanar , Lebanon
4 Reviva Regenerative Medicine Center, Middle East Institute of Health University Hospital , Bsalim , Lebanon
Background: Most common forms of hair loss (alopecia) are caused by aberrant hair follicle cycling and changes in hair follicle morphology. However, current treatments for alopecia do not specifically target these processes. Adiposederived stromal vascular cells (ADSVCs) that can be harvested from fat cells are one of the latest breakthroughs in the aesthetic field. The potential use of stem cell-based therapies (SCBT) for the repair and regeneration of various tissues and organs offers a paradigm shift that may provide alternative therapeutic solutions, which can be applied to prevent hair loss. This study aimed to present clinical cases of SCBT for the treatment of alopecia areata by transplantation of ADSVCs in the scalp. Methods: Twenty patients (9 women and 11 men) were recruited to our retrospectively registered study. After lipoaspiration, autologous ADSVCs were generated and characterized before the injection of 4-4.7 × 106 cells into the scalp of the patient. Hair regeneration was assessed by three clinical tests: the pull test, hair quality, and hair density. Results: All patients experienced hair regeneration, increased hair growth and decreased pull test 3 and 6 months after the treatment with ADSVCs [hair density (85.1 ± 8.7 vs 121.1 ± 12.5 hair/cm2, P < 0.0001), hair diameter (60.5 ± 1.8 vs 80. 8 ± 2.4μ, P < 0.0001) and pull-test values (4.4 ± 0.3 vs 0.8 ± 0.2, P < 0.0001), untreated versus 6 months post-operative)]. Significant variation was observed between men and women only for hair diameter. No significant differences were observed with age. Conclusions: The obtained results prove the efficacy and the safety of the treatment, and satisfaction of the patients confirm the quality of the results.
Baldness; Adipose-derived stromal vascular cells; Hair fall treatment; Stem cell; Mesenchymal stem cell; Hair loss; Alopecia
Hair loss is one of the most common complaints among
all patients consulting a dermatologist and is usually
associated with severe psychological disturbances, distress,
and symptoms of depression. Most common forms of
hair loss (alopecia) are caused by aberrant hair follicle
cycling and changes in hair follicle morphology. Cells
with stem cell properties have recently been described in
many integument appendages including feathers and
teeth, but the hair follicle stands out as one of the best
model systems for studying adult stem cells [
follicles are accessible, well defined in terms of their
developmental biology, and their stem cell populations are
located in discrete compartments or niches [
Among several factors, alterations in hair follicle size
may affect the hair loss: in fact, the size of a hair follicle
is dependent of the volume of its dermal papilla which
depends on the number of cells it contains [
There are three phases of hair growth, which every
hair follicle undergoes. The first phase is when follicles
undergo extremely rapid epithelial cell division and
execute exquisitely timed differentiation programs when in
the growing (Anagen) phase. The second phase is when
follicles growth stops during a certain period, the
(Catagen) phase [
]. The third phase is when most
follicles regress into structures that resemble immature
developing follicles, after which they go into a period of
mitotic quiescence, the (Telogen) phase. Stem cells are
at the core of all hair dynamic events that includes a
new burst of activity and further morphogenetic
remodeling as the follicle grows again at the start of a new
Anagen phase [
The term “stem cell” is given to a cell which has the
ability to self-renew as well as to differentiate into defined
cellular subtypes. Multipotent stem cells are present in
different adult tissues such as bone marrow, dental pulp,
adipose tissue, etc. [
]. Multipotent stem cells within
adipose tissue [
], existing in adipose-derived stromal
vascular cells (ADSVCs), are one of the most promising stem
cell population identified, since human adipose tissue is
easily obtained in large quantities with little patient
discomfort and secretory factors from ADSVCs have been
considered as a promising therapy for skin aging [
Therefore, the use of autologous ADSVCs can be
promising for hair loss. Since the stromal vascular fraction (SVF)
is saturated with stem cells among other cells derived
from adipose tissue, cells can be called ADSVFC if they
are used freshly, or ADSC/ADASC or others terms in
cases of primary cells placed in culture having then the
adherent feature, resulting in a set of mesenchymal stem
cells (MSCs). In fact, contrary to cultured ADSCs, freshly
isolated ADSVCs were shown to be highly positive for
CD34, and positive for CD117 and HLA-DR. MSCs
derived from adipose tissue when obtained by culture are
mostly negative for CD34, and HLA-DR. This indicates
clearly that primary cells are significantly more promising
in case we need to maintain a certain level of CD34 in the
In the present study, we aimed to use autologous
ADSVCs graft for the treatment of alopecia areata and
to assess the safety and effectiveness of the
transplantation. The clinical trial of 20 patients shows the use of
ADSVCs for hair growth and improvement as a valuable
White healthy subjects (n = 20, 38.3 ± 2.3 years, 9 women
and 11 men) from the Middle East were enrolled in the
study with no notable pathologic history in particular of
hair diseases, with confirmed diagnostic of hair loss
alopecia areata and all the selected subjects showed partial
alopecia grade 1 or 2 at Ludwig Scale [
]. Subjects were
excluded from this study if they had: histories of hair
diseases other than hair loss/alopecia areata, conditions
including moderate or severe head injury, burns, skin
diseases, stroke, cerebral or bone damage particularly of
the scalp or malignancies, brain abnormalities, learning
disability, major medical or psychiatric illness in the
previous 6 months, any metabolic/cardiovascular disease or
evidence of cardiac/renal damage or malignancies, diabetes,
hypertension, alcohol use, smoking, loss of weight during
the last 2 years, chemotherapy, immunosuppressive, head/
brain/abdominal surgeries, hormonal imbalances due to
one or multiple factors (such as menopause, stress,
depression, postpartum, chemotherapy, birth control, thyroid
disorders, ovarian cysts, or others), medications known to
cause hair loss (among others, medications for blood
pressure, heart diseases, contraception, depression, etc.),
disease and illness that may cause hair loss, chemical hair
treatments, traction alopecia, compulsive hair pulling
(Trichotillomania), poor nutrition, local infection (such as
hepatitis, syphilis, herpes, HIV, etc.) or allergic reaction,
abnormal physiologic levels of a comprehensive metabolic
panel (CBC), vitamins, lipid and liver panel, hematocrit,
hemoglobin, iron, ferritin, creatinine, coagulation factors,
C-reactive protein, erythrocyte sedimentation rate,
antinuclear antibodies, thyroid hormones, free and total
testosterone, follicule-stimulating and luteinizing hormone. Treated
alopecia areata cases were excluded; some example of
treatments (steroid injections, corticosteroid creams and
ointments, photochemotherapy, aromatherapy,
acupuncture, herbal supplements, vitamins, platelet-rich plasma
treatment, or any medications such as statins or other
Annexin V(+)/PI(+)/7AAD(+) (late apoptotic, necrotic cells) 0.17 ± 0.82
2 × 105 cells were labeled with fluorescence-coupled antibodies against the
indicated cell surface markers, Annexin V, 7AAD, propidium iodide solution (PI),
and analyzed using a MACSQuant flow analyzer as indicated in Materials and
Methods. The results are expressed as the mean ± SEM of all subjects (n = 20)
each performed in duplicate
ADSVC adipose-derived stromal vascular cell, SVF stromal vascular fraction
% of total cells
71.06 ± 5.08
10.28 ± 4.11
32.44 ± 2.09
18.56 ± 2.83
71.27 ± 3.54
27.70 ± 6.33
27.07 ± 5.81
55.27 ± 4.16
6.37 ± 4.12
18.62 ± 4.92
24.45 ± 2.17
3.19 ± 2.05
98.04 ± 1.12
10.17 ± 3.62
98.97 ± 0.71
1.02 ± 0.54
plasma cholesterol drug treatment, JAK-STAT pathway
inhibitors, plaquenil, antihypertensive vasodilator medication
(minoxidil), anthralin, immunomodulator therapy with
squaric acid dibutylester (SADBE),
diphenylcyclopropenone (DPCP) or the 5α-reductase inhibitor (finasteride),
home remedies or others). Blood samples and abdominal
fat lipoaspirates were collected from all the participants.
No previous treatments were given to the patient before
Three clinical tests, namely the pull test, hair quality, and
hair density were performed, and pictures were obtained
before and 3 and 6 months after the procedure to assure
the authenticity of the results. The pictures were taken in
the following positions to show the quantity of hair per
square centimeter: from the front, of the parietal scalp,
and close up [
The lipoaspirate [
] was diluted with sterile
phosphatebuffered saline (PBS), Sigma-Aldrich, St. Louis, MO, USA)
supplemented with antibiotics and centrifuged at 430 × g
for 10 min (without brakes) to remove contaminating
debris and red blood cells. The wash step was repeated 2–3
times depending on the quality of the specimen. The
floating adipose tissue was digested with an equal volume of
collagenase type I [10 mg/mL in PBS containing 5 mM
Ca2+/Mg2+ (C0130, Sigma-Aldrich), final concentration 0.
5%] at 37 °C for 30 min with shaking (250 rpm). The
collagenase was inactivated by adding an equal volume of
autologous serum, and the sample was centrifuged at 600 ×
g for 10 min. After centrifugation, the supernatant was
discarded and the cell pellet was resuspended in NaCl 0.
9% (Alpha Laboratories, Eastleigh, UK) and filtered
through a 100 μm cell strainer (CS003 – PNC
International Co. Ltd., Seoul, Korea) to remove debris. After
centrifugation (300 RCF/5 min), 5 ml of the stromal
vascular fraction were collected. All the processing must be
realized within a maximal time of 90 min. The number of
viable cells were determined manually (Trypan blue
method) and validated on MACSQuant analyzer
(Miltenyi-Biotech, Bergisch Gladbach, Germany) (7AAD
staining method). All the quality control tests and injections
were done with the obtained fresh samples.
Diameter: fine hair ≤ 60 μ; medium hair 60 to 80 μ, good hair ≥ 80 μ; Density: quantity of hair per cm2, normal is 175 to 300 hair/cm2; Pull test is the quantity of
hair pulled with the finger pull test, normal is zero. 0.2 ml per spot of injection was delivered perpendicularly, separated by 1 cm in a square shape all over the
scalp and a total of 5 ml were injected in 25 spots. Cell viability and apoptotic index as assessed by labeled cells with Annexin V/propidium iodide (PI)/7AAD, are
expressed respectively as the percentage of Annexin V(−)/PI(−)/7AAD(−) or Annexin V(+) cells divided by total cells
ADSVCs adipose-derived stromal vascular cells
Assessments of cell immunophenotyping, viability, apoptosis, and telomerase activity
Freshly isolated cells were characterized for ADSVCs
surface protein expression [
] by flow cytometry
(MACSQuant analyzer, Miltenyi-Biotech) according to
the manufacturer’s instructions. Cells were stained with
the following antihuman-conjugated monoclonal
antibodies: CD13-APC, CD14-PE, CD29-FITC, CD31-APC,
CD34-PE, CD45-VioBlue, CD73-APC, CD90-FITC,
CD105-VioBlue, CD144 (VE-Cadherin)-PE,
CD146Biotin, CD166-Biotin, HLA-ABC-FITC,
HLA-DRVioBlue, or relevant isotype-matched controls
(MiltenyiBiotech). Isotypes controls and automated compensation
were settled to minimize false positive fluorescence and
spectral overlap of fluorochromes respectively. Cell
viability and apoptosis were assessed by the
7AAD/AnnexinV/PI assay. In fact, cell viability was first assessed
manually using the Trypan blue (Sigma-Aldrich)
exclusion assay, and the results were then validated by the
7AAD method by flow cytometry. The telomerase
] was assessed by real-time qPCR (LightCycler 2.0,
Roche, Basel, Switzerland) using the Quantitative
Telomerase Detection Kit (Cat#MT3012, Allied Biotech,
Taipei, Taiwan) according to the manufacturer’s
ADSVCs injection and patient follow-up
The ADSVCs were injected into the scalp of the patient
according to the following procedure: (1) the upper frontal,
biparietal, and upper pyramidal area were first treated with
the aseptic chlorhexidine without local anesthesia; (2) to
reach the hair follicle area, the injection into the scalp area
was performed with the following attributes: syringe,
1 cm3; needle, 4 mm; gauge, 30; and depth, 4 mm [
0.2 ml per injection was delivered perpendicularly,
separated by 1 cm in a square shape all over the scalp
marked previously; a total of 5 ml were injected in 25
spots; (4) after the injection was administered, the needle
was kept in the scalp for 2 s. After the transplantation, the
patient was prescribed nonsteroid anti-inflammatory and
cephalexin antibiotics for 3 days. Patients were advised not
to shower until 24 h after the procedure, not to sunbathe
until after 1 week, and not to engage in sports until after
1 week; however, return to work can be on the same day.
Follow-up for hair evaluationwas based on the hair cycles
Fig. 1 Difference in hair diameter between patients before and 3 or
6 months after ADSVCs transplantation. a Whole population.
b Variation between male and female. ***P < 0.0001
and was performed 1 week, 3 months, and 6 months after
For descriptive statistics, mean and variance were
reported when appropriate. SPSS version 21.0 (IBM Corp.,
Armonk, NY, USA) was used for the statistical analysis.
Comparisons between groups were performed using
one-way ANOVA. Chi-square tests were applied to
detect the difference in the rates between different groups.
P values of less than 0.05 were considered significant.
In this study, the effect of hair regenerative ADSVC
therapy was evaluated in 20 patients (9 females and 11
males) aged between 23 and 63 years old. First, based
on a joint statement of the International Federation for
Adipose Therapeutics and Science (IFATS) and the
International Society for Cellular Therapy (ISCT)
published in 2013, which point out the minimal phenotypic
criteria to characterize the uncultured SVF population
from adipose tissue [
], we characterized these freshly
isolated cells. In fact, the immunophenotyping of the
transplanted cells showed clearly a heterogeneous
population of freshly isolated cells, which expressed not
only the mesenchymal stem cells markers, but also the
pericyte markers and particularly high levels of CD34.
These cells strongly expressed HLA-ABC but weakly
expressed HLA-DR markers (Table 1). Cell viability, as
assessed by Trypan blue and validated by 7AAD
staining, was > 96% and cells affected by an early apoptosis
were rare (Table 1); it is important to note that the total
time of processing was less than 120 min, and a
prolonged processing time affected the viability of the cells.
That is why, we managed the processing of all samples
during a maximal time of 120 min and transplanted the
cells in a total time not exceeding 3 h. A significant
decrease in the viability was observed after 4–6 h (8%,
24%, and 31% of decrement in cell viability rate after 4,
3H0a0ir hdaeinr/scimty2or trichometry is the quantity of hair per cm2, normal is 175 to
ADSVCs adipose-derived stromal vascular cells
5, and 6 h, respectively) (data not shown). On the other
hand, a total number of 4 to 4.7 × 106 cells were
transplanted: in fact, 0.2 mL containing 0.160–0.188 ×
106 cells were injected per spot (total = 25 spots, 5 mL).
In agreement with Varma et al. [
], our results
indicated that freshly isolated ADSVCs cells were
shown to be highly positive for CD34, contrary to the
expression of CD105 and especially CD166 which were
relatively low (3.19% and 6.37%, respectively) (Table 1):
to maintain a minimum level of these cells in the
sample, this led us to consider at least the presence of
5000 CD105+/CD166+ cells in the 0.2 mL of
transplanted sample per spot of injection, which
prompted us to choose the minimum concentration of
160,000 cells/spot of injection (= 4 × 106 total cells/25
spots/per subject). Importantly, to avoid any
aggregation of the cells, which was observed in cases
where the cell concentration was > 200,000 cells per 0.
2 mL, and to maintain the minimum levels of CD105+
and CD166+ cells per injection, a total of 4.0–4.7 × 106
cells was delivered to the subjects.
Second, we assessed the hair loss and growth which
were determined as changes in hair density (n/cm2) and
hair diameter (μ), as well as the pull test (Table 2).
Overall, 55% of the patients showed medium diameter
hair and 45% showed fine hair. All the study subjects
showed abnormal hair density (density < 175 hair/ cm2
in 100% of the subjects). All the patients showed a
value superior to 0 for the pull test. In addition, no
significant variations were observed with age.
ADSVCs injection increases hair diameter
Our results are based on the density of hair per square
centimeter, the diameter of the hair, and the pull test
performed manually by the same physician in all phases.
Table 3 shows the results obtained at 3 and 6 months after
the injection of ADSVCs in comparison to the preoperative
ones. As shown in Table 2, the hair diameter increased
significantly (P < 0.0001) with the injection of ADSVCs,
especially 6 months after the treatment (80.8 ± 2.4 μ and 62.8 ±
1.7 μ vs. 60.5 ± 1.8 μ for 6 and 3 months postoperatively vs.
preoperatively) (Fig. 1a). In total, 19 out of 20 patients
showed improved hair diameter; only one patient did not
show any improvement. Approximately, on average, 32%
improvement was obtained, and maximum improvement
was approximately 50.1%. Significant variation was
observed between males and females (Fig. 1b).
ADSVCs injection improves hair density
We can note that the hair density was significantly (P < 0.
0001) augmented after treatment with ADSVCs (121.1 ±
12.5 and 120.8 ± 12.6 vs. 85.1 ± 8.7 for 6 and 3 months
postoperatively vs. preoperatively) (Table 4). The mean
growth was approximately 36%, and the optimal effect
was 61.2% (Fig. 2a). The hair growth occurred during the
first phase. Of the 20 patients studied, only two did not
show any significant improvement. No significant
differences were observed between males and females (Fig. 2b).
ADSVC injection increases pull test
The results of the pull test showed a significant decrease
in the number of extracted hair (P < 0.0001) following
ADSVC treatments (0.80 ± 0.17 and 0.90 ± 0.20 vs. 4.35
± 0.33 for 6 and 3 months postoperatively vs.
preoperatively). We could notice that the hair became stronger at
3 and 6 months postoperatively, leading to values of
mainly 0 and 1 in the pull test (Table 5, Fig. 3a). In fact,
the values of the pull test in the control group ranged
between 2 and 8; however, they were markedly inferior
in the ADSVC-treated group. In total, 2 of the 20
patients showed no significant improvements. All other
patients had normal responses. In addition, no
significant differences were observed between males and
females (Fig. 3b).
Hair fall treatment is a challenge for many doctors;
however, hair fall treatment can be performed in
multiple ways. To date, no studies have shown the hair
fall treatment can be performed using ADSVCs with
satisfactory results. Our research is unique and the
first in the field of hair fall treatment to use ADSVCs.
Most of the studies conducted in the past were based
on hematopoietically derived plasma or
ADSVCconditioned medium (ADSVC-CM).
This study showed that the transplantation of
autologous ADSVCs is safe and effective and can be
considered an encouraging cell-based therapy for the treatment
of alopecia and a nonsurgical hair loss treatment. Hair
growth and thickness were markedly improved 6 months
after the treatment (Fig. 4).
Reports of the use of stem cells in the treatment
of alopecia are rare. Researchers have utilized bone
marrow/cord blood-derived stem cells or
ADSVCCM. However, to our knowledge, there is no study
that investigated the effect of ADSVCs for the
treatment of alopecia. Our results on the characterization
of the isolated and transplanted cells agreed with the
previously described immunophenotype [
17, 21, 22
Li et al. [
] recently showed that patients with
severe alopecia areata showed improved hair regrowth
and quality of life after receiving stem cell educator
therapy. They demonstrated the safety and efficacy
of a new method where the mononuclear cells are
separated from the whole blood and were allowed to
briefly interact with adherent human cord
bloodderived multipotent stem cells, and the “educated”
autologous cells were returned to the patient’s
circulation. Fukuoka et al. [
] showed that treatment
with ADSVC-CM effectively activated hair
regeneration; ADSVC-CM is rich in growth factors such as
vascular endothelial growth factor, hepatocyte growth
factor, platelet-derived growth factor, and insulin-like
growth factor 1. Another study with a female pattern
hair loss treated with ADSVC-CM showed that the
treatment increased the hair density and thickness
]. Recently, it was reported that autologous bone
marrow-derived mononuclear cells seem to be a safe,
tolerable, and effective treatment for the
management of both resistant alopecia areata and
androgenetic alopecia [
The injected ADSVCs in this study might release
growth factors, thus promoting vascularization,
encouraging new capillaries to form, increasing the
production of hair, and improving the supply of blood to
the scalp. This provides an ideal environment for the
hair follicles to grow new, denser, and healthy hair. This
research is the beginning of many series of studies that
will be conducted in the future to improve the results
and decrease the cost of the procedure to make it more
efficient and affordable to patients. It is important to
note that most cases of alopecia areata (approximately
80%) resolve spontaneously especially first cases of
alopecia areata. It will be interesting to study the efficacy
of ADSVCs transplantation for grade III and advanced
cases of alopecia areata.
In conclusion, treatment using ADSVCs appears highly
effective for alopecia areata and may represent a new
avenue of therapy for hair regeneration. ADSVC
injection promotes good stability of the hair by increasing
the hair density, the hair diameter, and decreasing the
pull test to almost zero. Furthermore, patients must be
very well selected depending on their lifestyle, the cause
of hair fall and baldness grade to obtain a good result
with this procedure.
ADASC: Adipose-derived adult stem cell; ADSC: Adipose-derived stem cell;
ADMSC: Adipose-derived-mesenchymal stem cell; ADSVC: Adipose-derived
stromal vascular cell; ADSVC-CM: ADSVC-conditioned medium; CD: Cluster of
differentiation; IFATS: International Federation for Adipose Therapeutics and
Science; ISCT: International Society for Cellular Therapy; MSC: Mesenchymal
stem cell; PBS: Phosphate-buffered saline; SCBT: Stem cell-based therapies;
SVF: Stromal vascular fraction
The authors would like to acknowledge all parties that participated in this
Personal funding by RA.
Availability of data and materials
All data generated or analyzed during this study are included in this
published article and its additional files.
NM and RA contributed to conception, study design, and conduction of the
study. RA, NM, FR, and AH contributed to experimentation and data
collection. RA, NM, FR, and AH contributed to data analysis and
interpretation. RA, NM, and AH contributed to manuscript writing and
editing. RA and NM contributed to patient selection and sample
procurement. AA contributed to study management. All authors read and
approved the final manuscript.
Ethics approval and consent to participate
The study was conducted in strict adherence to the tenets of the
Declaration of Helsinki, and it was retrospectively registered in
ClinicalTrials.gov (Identifier: NCT03427905). The protocol was approved by
the institutional review board of REVIVA Regenerative Medicine Center at the
Middle East Institute of Health University Hospital as previously [
under the academic regulations of the public institution, the Lebanese
University. All patients provided written informed consent and fresh samples
were procured by Dr. Rami Andari (plastic surgeon) from REVIVA at the
Middle East Institute of Health (MEIH) University Hospital. The first patient
was enrolled in March 2013.
The authors declare that they have no competing interests related to this
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
1. Blanpain C , Lowry WE , Geoghegan A , Polak L , Fuchs E . Self-renewal, multipotency, and the existence of two cell populations within an epithelial stem cell niche . Cell . 2004 ; 118 ( 5 ): 635 - 48 .
2. Kim JY , Tavare S , Shibata D . Human hair genealogies and stem cell latency . BMC Biol . 2006 ; 4 : 2 .
3. Ohyama M , Terunuma A , Tock CL , et al. Characterization and isolation of stem cell-enriched human hair follicle bulge cells . J Clin Invest . 2006 ; 116 : 249 - 60 .
4. Morris RJ , Liu YP , Marles L , et al. Capturing and profiling adult hair follicle stem cells . Nat Biotech . 2004 ; 22 : 411 - 7 .
5. Elliott K , Stephenson TJ , Messenger AG . Differences in hair follicle dermal papilla volume are due to extracellular matrix volume and cell number: implications for the control of hair follicle size and androgen responses . J Invest Dermatol . 1999 ; 113 : 873 - 7 .
6. Yoo BY , Shin YH , Yoon HH , Seo YK , Park JK . Hair follicular cell/organ culture in tissue engineering and regenerative medicine . Biochem Eng J . 2010 ; 48 : 323 - 31 .
7. Panteleyev AA , Jahoda CA , Christiano AM . Hair follicle predetermination . J Cell Sci . 2001 ; 114 : 3419 - 3431 .
8. Alonso L , Fuchs E. The hair cycle . J Cell Sci . 2006 ; 119 : 391 - 3 .
9. Sun TT , Cotsarelis G , Lavker RM . Hair follicular stem cells: the bulgeactivation hypothesis . J Invest Dermatol . 1991 ; 96 : 77S - 8S .
10. Ludwig E. Classification of the types of androgenetic alopecia (common baldness) occurring in the female sex . Br J Dermatol . 1977 ; 97 : 247 - 54 .
11. Zuk PA , Zhu M , Ashjian P , et al. Human adipose tissue is a source of multipotent stem cells . Mol Biol Cell . 2002 ; 13 : 4279 - 95 .
12. Park BS , Jang KA , Sung JH , et al. Adipose-derived stem cells and their secretory factors as a promising therapy for skin aging . Dermatol Surg . 2008 ; 34 ( 10 ): 1323 - 6 .
13. Hillmann K , Blume-Peytavi U . Diagnosis of hair disorders . Semin Cutan Med Surg . 2009 ; 28 ( 1 ): 33 - 8 .
14. Van Neste D , Trüeb RM . Critical study of hair growth analysis with computerassisted methods . J Eur Acad Dermatol Venereol . 2006 ; 20 ( 5 ): 578 - 83 .
15. Jahoda CAB , Reynolds AJ . Dermal-epidermal interactions. Adult folliclederived cell populations and hair growth Dermatol Clin . 1996 ; 14 : 573 - 83 .
16. Li K , Gao J , Zhang Z , et al. Selection of donor site for fat grafting and cell isolation . Aesth Plast Surg . 2013 ; 37 : 153 - 158 .
17. Mitchell JB , McIntosh K , Zvonic S . Immunophenotype of human adiposederived cells: temporal changes in stromal associated markers . Stem Cells . 2006 ; 24 : 376 - 85 .
18. Vonderheide RH . Telomerase as a universal tumor-associated antigen for cancer immunotherapy . Oncogene . 2002 ; 21 : 674 - 9 .
19. Goodell GG , Gallagher FJ , Nicoll BK . Comparison of a controlled injection pressure system with a conventional technique . Oral Surg Oral Med Oral Pathol Oral Radiol Endod . 2000 ; 90 ( 1 ): 88 - 94 .
20. Bourin P , Bunnell BA , Casteilla L , et al. Stromal cells from the adipose tissuederived stromal vascular fraction and culture expanded adipose tissuederived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT) . Cytotherapy . 2013 ; 15 ( 6 ): 641 - 8 . https://doi.org/10. 1016/j.jcyt. 2013 . 02 .006.
21. Varma MJ , Breuls RG , Schouten TE , et al. Phenotypical and functional characterization of freshly isolated adipose tissue-derived stem cells . Stem Cells Dev . 2007 ; 16 ( 1 ): 91 - 104 .
22. Planat-Benard V , Silvestre JS , Cousin B , et al. Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives . Circulation . 2004 ; 109 : 656 - 63 .
23. Li Y , Yan B , Wang H , et al. Hair regrowth in alopecia areata patients following stem cell eEducator therapy . BMC Med . 2015 ; 13 : 87 .
24. Fukuoka H , Suga H . Hair regeneration treatment using adipose-derived stem cell conditioned medium: follow-up with trichograms . Eplasty . 2015 ; 15 : e10 .
25. Shin H , Ryu HH , Kwon O , Park BS , Jo SJ. Clinical use of conditioned media of adipose tissue-derived stem cells in female pattern hair loss: a retrospective case series study . Int J Dermatol . 2015 ; 54 ( 6 ): 730 - 5 .
26. Ibrahim ZA , Elmaadawi IH , Mohamed BM , et al. Stem cell therapy as a novel therapeutic intervention for resistant cases of alopecia areata and androgenetic alopecia . J Dermatol Treat . 2016 ; 24 : 1 - 10 .
27. Alió Del Barrio JL , El Zarif M , de Miguel MP , et al. Cellular therapy with human autologous adipose-derived adult stem cells for advanced keratoconus . Cornea . 2017 ; 36 ( 8 ): 952 - 60 .
28. Alió Del Barrio JL , El Zarif M , Azaar A , et al. Corneal stroma enhancement with decellularized stromal laminas with or without stem cell recellularization for advanced keratoconus . Am J Ophthalmol . 2018 ; 186 : 47 - 58 .