Kasper C., van Griensven M., P?rtner R. (Eds.) Bioreactor Systems for Tissue Engineering II: Strategies for the Expansion and Directed Differentiation of Stem Cells
Подождите немного. Документ загружается.
C57Bl/6 mice to B6D2F1 mice irradiated beforehand and compared ASC-injection
0, 7, and 14 days vs 14, 21, and 28 days post implantation.
The remarkable finding was that the early implanted animals had significantly
higher survival rates due to GVHD than the later implanted animals.
In contrast to the just mild suppression of ASC-CM alone, Kang et al. found the
supernatant of ASC-CM derived from beagle dogs culture to suppress leukocyte
proliferation that was stimulated before with Concovalin A, pokeweed mitogen,
and LPS [142]. For more detailed consideration of the immunomodulatory cytokine
profile of these canine ASC, see the p aragraph above concerning cytokine profiles
of ASC.
In 2004, LeBlanc et al. reported a case of severe therapy-re sistant GVHD of the
gut and the liver, which improved rapidly after injection of ASC, which displayed
the first use of ASC in clinical medicine [173].
From 2006 on, Fang et al. published a series of studies dealing with the clini cal
application of ASC for the suppression of GVHD [46, 174–178]. In their study from
2006, they reported a case of a patient who suffered from severe GVHD after stem
cell transplantation, which also manifested mainly in the gut and the liver, proven
by coloscopy and elevated liver enzymes [178].
After ASC-transplantation from an allogenic donor, symptoms vanished in a few
days, indicating immunosuppression.
Another case report referred to a woman with an acute hepatic GVHD cause d
by hematopoietic s tem cell transplantation, who failed conventional immu-
nosuppressive therapy, e.g., cyclosporine and prednisone [177]. Thus, she was
treated with tacrolimus, which was discontinued because of deterioration of
renal function. As a salvage therapy, she was than treated with ASC, which
resulted in rapid and complete resolution of hepatic GVHD as well as renal
toxicity.
Similar finding were published, when two pediatric patients suffered from
Philadelphia chromosome-positive acute lymphatic leukemia (ALL) or acute mye-
loic leukemia (AML), respectively [176]. The child with ALL displayed a gastro-
intestinal manifestation with severe, partly bloody diarrhea, significant loss of
weight, and deterioration of general condition 89 days after hematopoietic cord
blood transplantation although adequate pharmaceutical immunosuppression was
administered.
AML in the other patient was treated with peripheral blood stem cell transplan-
tation and an adequate pharmaceutical therapy regime as well. After 62 days, a
strong involvement of the liver in GVHD was observed by elevation of the liver
enzymes.
Both patients showed rapid remission of symptoms after injection of allogenic
ASC without side effects, which lasted at least for the follow-up of 374 days or
2 years after ASC-injection, respectively.
Comparable success was noticed in six patients in another study suffering from
steroid-refractory GVHD after hematopoietic stem cell transplantation b ecause
of leukemia [175]. Here different regimes of immunosuppressive therapy were
also followed to protect the patients from GVHD, but their condition deteriorated.
Isolation, Characterization, Differentiation, and Application of Adipose-Derived Stem Cells 87
Following ASC transplantation, four of the patients kept remission-free for the
duration of the follow-up, which ranged between 18 and 90 months. Two of the
patients died – one with no obvious response to AMC transplantation of multi-
organ failure and one of a relapse of leukemia. However, the patient mentioned
finally showed a good response to ASC-therapy while the relapse was 16 month
after the ASC-infusion; thus the proven response rate can be stated as five of s ix
patients.
The two studies p ublished lately were case reports of the treatment of either
refractory pure red cell aplasia (PRCA) after major ABO-i ncompatible stem
cell transfusion or refractory chronic autoimmune thrombocytopenic purpura
[46, 174].
PRCA occurred in two patients in the first mentioned study due to donor-
recipient incompati bility, which here was HLA- but not ABO -matched. Although
a conditioning therapy with Busulfan and cyclophosphamide was performed in
both cases, the clinical appearance of PRCA, i.e., prin cipally reticulocytopenia
and thus erythrocytopenia, could not be avoided. Both patients needed a red
blood cell transfusion weekly with no beneficialeffectfromerythropoietin
administration.
In the second case, o ne patient was diagnosed with autoimmune thrombocyto-
penic purpura, a disorder wherein autoantibodies are directed agai nst platelet
surface glycoprotein (GPs), usually GPIIb/IIIa or GPIb/IX. Hereby, splenic platelet
destruction is induced and platelet production inhibited, leading to lack of platelets
and extended clotting time.
As in the previously described cases, ASC transplantation led to remission,
which is continuing up to the publishing date.
In four studies carried out by the work-group around Gonzalez, ASC were also
shown to suppress inflammative diseases [88, 89, 179, 180]. Two studies concerned
the treatment of inflammative bowel diseases, induced by tri-nitro-benzene sulfonic
acid, and sepsis, the two other studies concerned the treatment of an experimental
arthritis, induced by immunization against collagen. All of the studies were per-
formed in vitro as well as in vivo in a murine model.
The authors’ interesting thesis is that ASC intervene in the regulation of Th1-
cells, the key effectors of autoimmune disorders like Crohn’s disease or rheumatoid
arthritis, by influencing regulatory T-cells which can be considered as influential
mediators of regulating the inflammatory response.
Actually, they found downregulation of both Th1-driven autoimmune and
inflammatory responses, resulting in amelioration of the clinical and histopatholo-
gical severity, abrogating body weight loss, diarrhea, and overall survival in the
colitis/sepsis studies and joint inflammation in the arthritis studies.
These findings were med iated by suppression of Th1-cells as wells as augmen-
tation of IL-10-producing T-cells, a cytokine responsible for inhibition of T-cell-
response.
Also, ASC impaired Th1 cell expansion by direct cell-to-cell contacts and,
additionally induced a population of CD4(+)CD25(+)FoxP3(+) regulatory T cells
with suppressive capacity on Th1 effector responses in vitro and in vivo.
88 J.W. Kuhbier et al.
Taking these findings together, ASC have a great immunomodulatory influence
as they are effective suppressors of immune system responses, in particular T cell
responses which mediate GVHD as well as autoimmune inflammatory reactions. In
the mentioned pioneering clinical studies, they acted as salvage therapy for cases
that were refractory to conventional therapy.
Therefore, they may play a bigger role in future directions of immunologic and
hematologic therapy as well as in transplantation medicine due to their properties
with seemingly lack of side effects.
5.6 Other Purposes in Current In Vivo Application
A quite interesting clinical application for ASC was published by Garcia-Olmo
et al. in 2003 describing a case report, in which they used autologous ASC in a
fistula in Crohn’s disease [181]. Be side the characteristic diarrhea caused by
transmural inflammation of the intestine wall, ente rocutaneous, rectovaginal, and
perianal fistulas display a frequent and often stigmatizing complication of Crohn’s
disease with an incidence ranging between 17 and 50%, depending on the sourc e. In
particular, secreting processes, e.g., vaginal flatus or fetal incontinenc e in rectova-
ginal fistulas or secretion of intestinal content through enterocutaneous fistulas, are
extremely unpleasant for the patient.
In the reported case, the patient had a history of disease for 11 years prior to the
study and had already received several therapeutic trials including surgical treat-
ment, which resulted in remission.
Garcia-Olmo and coworker harvested the SVF via liposuction from subcutane-
ous fat, processed it, and injected purified ASC superficial into the wall of the
fistula. A vaginal flap covered the resected posterior vaginal wall while fibrin glue
sealed the perineal hole. After 1 week, the fistula had completely closed, demon-
strating epithelization. Due to this, a clinical phase I study was initiated including
five patients suffering from Crohn’s fistulas that were also treated at least twice
medically and surgically [182]. Here, in 75% of the cases, a closure and epitheli-
zation of the fistula, stated as healing, occurred. Nevertheless, it is worth mention-
ing that they found no relationship between the numbers of cells injected and the
success of the procedure as well as between the age or gender of the patient and the
success.
Two other studies followed, the first being a clinical phase II trial including 35
patients suffering from complex perianal fistulas, and other than Crohn’s fistulas,
which resulted as well in a healing in 71% of the patients [183]. In the second, the
effectiveness of injection of either the SVF or purified and expanded ASC was
tested with much better results for the expanded ASC, i.e., healing in 75% of the
fistulas vs 25% of the fistulas with the SVF. Obviously, this is due to the content of
just 2–5% ASC in the SVF witho ut processing [184 ].
The same work-group around Garcia-Olmo also used a similar technique for
successful treatment of a tracheomediastinal fistula [185]. The patient had
Isolation, Characterization, Differentiation, and Application of Adipose-Derived Stem Cells 89
received treatment with Nd-YAG laser resulting in progressive necrosis which led
to formation of a fistula with a diameter of 10 mm. Within 3 h of general
anesthesia, a lipoaspirate was harvested, centrifuged, purified, and resuspended
infibringluefollowedbyabronchoscopyinwhichthefibringluewiththeASC
was injected into the cavity of the fistula. General examination concerning
clinical symptoms was performed weekly for the first month; flexible bronchos-
copywasdoneevery3months,whichrevealedlackofsymptomsaswellas
mayor reepithelization and complete closure of the fistula, leaving only a small
depression in the tracheal wall.
Another application already used in clinical medicine is the so-called cell-
assisted lipotransfer by the work-group around Yoshimura [186–188], though the
first description of implantation of ASC for soft tissue augmentation was by Cho
et al. [139]. Either in ASC pretreated in conditioned medium or undifferentiated
ASC were implanted subcutaneously in athymic mice, each group supplem ented
with or without bFGF. Six weeks after implantation, they found not just newly
formed adipose tissue but also neovascularisation of this tissue with both prediffer-
entiation and bFGF enhanced these effects.
These finding were approved by Mat sumoto et al. who used merely centrifu-
gation to yield human SVF, which was m ixed 1:1 w ith aspirated f at to obtain
fat wit h a high ASC con cen tra tion (cell-assisted lipotransfer; CAL), labeled with
CM.DiI and transplanted it also subcutaneously to severe combined immune
deficiency mice [186]. The CAL fat survival was better than the pure fat controls,
resulting in 35% larger specimen derived a fter transplantation, with SVF-cells
stained DiI-positive found between adipocytes and connective tissue. Some of
these cells were also positive for von Will ebrand factor, suggesting endothelial
differentiation.
CAL transplantation in Green fluorescent protein rats also showed neoangiogen-
esis by SVF-cells in the acute phase of transplantation, indicating CAL as
promising tool for long-time survival of fat transplantation.
Actually, this work-group performed two clini cal trials, one dealing with facial
lipoatrophy and one with breast augmentation, which could prove the success of
CAL [187, 188].
In the lipoatrophy treatment study, six patients received autologous SVF from
the abdomen which resulted in an average volumetric augmentation between 60 and
80% in the 10 month follow-up examination [188]. As facial lipoatrophy is very
stigmatizing because patients can hardly hide it, those improvements display a
satisfying result, though refinements should always be aimed for.
The second study examined breast augmentation by CAL in 40 cases. Although,
the technique disp layed long-term survival and safety, two cases with small cyst
formation and microcalcification were detected by mammography at 24 months
[187]. Less fat atrophy was observed and the augmented breasts looked more
natural than those with artificial implants, which also generally appea red to be
harder than the CAL-augmented ones.
Although those findings are impressive, they still raise the question whether
CAL is really superior to (pure) fat transfer by methods of proven quality and by an
90 J.W. Kuhbier et al.
experienced surgeon. With a careful, gentle, and tissue-conserving technique dur-
ing liposuction, comparable and repeatable results can usually be achieved, as was
reviewed recently in the literature [67, 189].
6 Tissue Engineering Applications
As a general definition, tissue engineering combines the principles of bioengineer-
ing, biomaterial engineering, and cell transplantations to generate bio-artificial
tissues and organs, thus stimulating the self-regeneration of damaged tissues after
the in vivo transfer. Artificial ECM is provided to allow for cell proliferation and
differentiation in a three-dimensional cell scaffold. Generally, two classes of
biomaterials are used for tissue engineering purposes naturally derived and
synthetic materials; the decision usually depends on the physiological and mechan-
ical requirements as well as the biodegradability.
6.1 Adipose Tissue
In an early appro ach, Patrick et al. seeded preadipocytes on poly DL-lactic-co-
glycolic acid (PLGA) scaffolds where they differentiated into mature adipocytes,
although they did not reach the size of natural mature adipocytes isolated from
epididymal adipose tissue [190].
Adipogenic differentiated rat ASC were seeded on polyglycolic acid (PGA) fibers
and implanted subcutaneously onto the heads of rats. Intracellular lipid vacuoles
could be demonstrated histologically in the grafts explanted after 4 and 8 weeks [191].
In the study of von Heimburg et al., human preadipocytes were seeded on
different biodegradable carriers like HYAFF 11 sponge and collagen sponges.
The HYAFF 11 sponges turned out to have a higher cell density than the collagen
matrices used [192]. Further modifications including pore size and coating with
glycosaminoglycan hyaluronic acid improved cell penetration and vascularization
in an in vivo appro ach [193]. Coll agen sponges were also used by Huss et al. for
coculture assay with human mammary epithelial cells and preadipocytes. They
found lipid-containing cells clustered around ductal structures [194].
Gelatin sponges seeded with human ASC have soft tissue-like mechanical pro-
perties which is generally preferable compared to more rigid structures [195].
Hyaluronic acid (HA)-based materia ls have been used in a variety of tissue engi-
neering purposes and clinical applications [196, 197]. An HA-based scaffold has
also been used as a scaffold for adipocyte precursor cells. Full maturation into
adipocytes was achieved with an even cell distribution [198].
In a unique approach, in 2008 Valle
´
e et al. [199] completely renounced the
substitution of exogenous matrix materials, resulting in three-dimensional cell
sheets of adipogenic stimulated cells.
Isolation, Characterization, Differentiation, and Application of Adipose-Derived Stem Cells 91
6.2 Bone Tissue
Three dimensional culture has a favorable effect on expression of osteogenic marker
genes compared to monolayer culture when ASC are cultured in osteogenic
medium [200]. The use of stromal vascular fractions of adipose tissue has been
considered advantageous since it enables direct three-dimensional seeding without
two-dimensional culture ste ps for cell expansion [201]. ASC seeded on biomater-
ials of clinical relevance in the treatment of bone defects like hydroxyapatite,
cancellous human bone fragments, deproteinized bovine bone granules, and tita-
nium produce more calcified matrix than cells in monolayers [202].
After 2 weeks of differentiation in porous PLGA foams, mineralized nodular
structures were shown [203]. In their rat transplantation model, Lee et al. demon-
strated that prediffer entiated grafts did not show bone formation but stained posi-
tively for osteocalcin after 4 weeks [191]. After 8 weeks, osteocalcin expression
was still detectable and, additionally, bone formation was shown in histological
analysis.
In their study with electrospun composite scaffolds consisting of beta-tricalcium
phosphate (TCP) crystals and poly
L-lactic acid (PLA), McCul len et al. provided
another indication that matrix properties are important in the differentiation process
as alkaline phosphatase activity and mineralization were shown to depend on fiber
diameter and TCP content [204].
Shen et al. used a three-dim ensional sintered microsphere matrix of poly(lactide-
co-glycolide) and differentiated using recombinant GDF-5 as an alternative matrix
material for bone tissue engineering [205].
Combinations of ECM components are often used to enhance the cell adhesion
on hydrophobic synthetic polymers [206].
Beside the administration of modifying substances like growth factors or the
transduction with genes of the osteogenic signaling pathway, an optimized dif-
ferentiation of the ASC is largely influenced by the used scaffold material and
stimulation exerted by mechanical forces. Honeycomb-shaped atellocollagen
sponges seeded with ASC were successfully used for bone formation in 3D
cultures in vitro and in vivo [207]. Osteoinductive materials like b-TCP have
been shown to induce osteogenic differentiation [208]. In a comparative study
between akermanite ceramics and b-TCP an even higher osteoinductivity was
found for the akermanite ceramics [209]. Peterson et al. demonstrated in a
combined approach that BMP-2 transfected ASC together with collagen-ceramic
carriers resulted in healing of a critical size femoral defect when applied to nude
mice [162].
In their approach dealing with the induction of osteogenesis by mechanical
forces, van Griensven et al. showed that bone tissue constructs were achieved by
seeding adult stem cells derived from bone marrow and adipose tissue seeded on
Sponceram matrices in a rotating bed bioreactor [210].
In a combined assay with human ASC seeded on a composite BMP-2 loaded
PLGA/hydoxyapatite scaffold were implanted subcutaneously on the backs of
92 J.W. Kuhbier et al.
athymic mice without any predifferentiation. After explantation, expression of
human-specific osteoblastic genes was found [211].
6.3 Cartilage Tissue
PLGA is a scaffold commonly used in cartilage tissue engineering based on
differentiated ASC [212–214]. In their study, Jin et al. combined cellular tranduc-
tion with human TGF b2 and cultivation on a three-dimensional PLGA/alginate
compound to produce cartilage formation efficiently in vitro and in vivo [215].
While copolymers of PLGA is a common substrate in cartilage tissue engineering
due to advantageous mechanical properties, successful cell seeding is often ham-
pered by its hydrophobicity [216]; the authors circumvented this problem by
choosing aliginate as a cell carrier.
In another approach ASC were encapsulated in aliginate microbeads together with
a chimeric RGD-protein. The parameters analyzed included chondrogenic differen-
tiation after TGFb3 dependent induction. An increased expression of chondrogenic
genes was observed which dependent on a b1 integrin mediated signaling [217].
Cheng et al. worked without stimulation with exogenous growth factors when
they seeded ASC on scaffolds derived from porcine articular cartilage. After a
cultivation period between 4 and 6 weeks, expression of chondrogenic genes was
observed [218]. These finding stress the importance of three-dimensional structures
in the chondrogenic differentiation for which there is a broad consensus in the
published literature [132–134, 217, 218]. Also mechanical forces like hydrostatic
pressure have been found to be important for chondrogenesis and have been implied
by Ogawa et al. in their study on ASC seeded in three-dimensional collagen
matrices. Cyclic hydrostatic pressure conditions caused an increase in chondro-
genic gene expression in TGFb1 stimulated cultures.
Another possibility for cartilage tissue engineering was described by Hildner
et al. when they demonstrated that cocultivat ion of human articular chondrocytes
together with ASC resulted in increased collagen type IX expression, indicating a
long-term stability of cartilage [219].
7 Summary
Stem cells derived from adipose tissue represent an enormous potential for medical
therapeutic applications. Their versatile properties range from promotion of cell
proliferation, differ entiation, and migration over immunomodulatory functions,
e.g., in GVHD and regeneration to facilitate tissue repair and neovascularization
in injury and ischemia. A major advantage for clinical application of ASC is based
upon easy harvesting techniques with sufficient yields, vast availability, and low
donor site morbidity compared to stem cells obtained from bone marrow or
Isolation, Characterization, Differentiation, and Application of Adipose-Derived Stem Cells 93
peripheral blood. The fast expanding field of basic and clinical research in ASC
biology, pathophysiology, and their potential therapeutic applications promises
further exciting discoveries in the near future.
References
1. Beltrami AP, Barlucchi L, Torella D, Baker M, Limana F, Chimenti S, Kasahara H, Rota M,
Musso E, Urbanek K, Leri A, Kajstura J, Nadal-Ginard B, Anversa P (2003) Adult cardiac
stem cells are multipotent and support myocardial regeneration. Cell 114:763–776
2. van VP S, JP DPA, Goumans MJ (2007) Isolation and expansion of resident cardiac
progenitor cells. Expert Rev Cardiovasc Ther 5:33–43
3. Dhawan J, Rando TA (2005) Stem cells in postnatal myogenesis: molecular mechanisms of
satellite cell quiescence, activation and replenishment. Trends Cell Biol 15:666–673
4. Peault B, Rudnicki M, Torrente Y, Cossu G, Tremblay JP, Partridge T, Gussoni E, Kunkel
LM, Huard J (2007) Stem and progenitor cells in skeletal muscle development, maintenance,
and therapy. Mol Ther 15:867–877
5. Griffiths MJ, Bonnet D, Janes SM (2005) Stem cells of the alveolar epithelium. Lancet
366:249–260
6. Kim CF, Jackson EL, Woolfenden AE, Lawrence S, Babar I, Vogel S, Crowley D, Bronson
RT, Jacks T (2005) Identification of bronchioalveolar stem cells in normal lung and lung
cancer. Cell 121:823–835
7. Brittan M, Wright NA (2002) Gastrointestinal stem cells. J Pathol 197:492–509
8. Bussolati B, Bruno S, Grange C, Buttiglieri S, Deregibus MC, Cantino D, Camussi G (2005)
Isolation of renal progenitor cells from adult human kidney. Am J Pathol 166:545–555
9. Herrera MB, Bruno S, Buttiglieri S, Tetta C, Gatti S, Deregibus MC, Bussolati B, Camussi G
(2006) Isolation and characterization of a stem cell population from adult human liver. Stem
Cells 24:2840–2850
10. Koblas T, Zacharovova K, Berkova Z, Mindlova M, Girman P, Dovolilova E, Karasova L,
Saudek F (2007) Isolation and characterization of human CXCR4-positive pancreatic cells.
Folia Biol (Praha) 53:13–22
11. Levy V, Lindon C, Zheng Y, Harfe BD, Morgan BA (2007) Epidermal stem cells arise from
the hair follicle after wounding. FASEB J 21:1358–1366
12. Nakatomi H, Kuriu T, Okabe S, Yamamoto S, Hatano O, Kawahara N, Tamura A, Kirino T,
Nakafuku M (2002) Regeneration of hippocampal pyramidal neurons after ischemic brain
injury by recruitment of endogenous neural progenitors. Cell 110:429–441
13. Ourednik J, Ourednik V, Lynch WP, Schachner M, Snyder EY (2002) Neural stem
cells display an inherent mechanism for rescuing dysfunctional neurons. Nat Biotechnol
20:1103–1110
14. Mimeault M, Hauke R, Batra SK (2007) Stem cells: a revolution in therapeutics-recent
advances in stem cell biology and their therapeutic applications in regenerative medicine and
cancer therapies. Clin Pharmacol Ther 82:252–264
15. Schaffler A, Buchler C (2007) Concise review: adipose tissue-derived stromal cells–basic
and clinical implications for novel cell-based therapies. Stem Cells 25:818–827
16. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick
MH (2001) Multilineage cells from human adipose tissue: implications for cell-based
therapies. Tissue Eng 7:211–228
17. Fruebis J, Tsao TS, Javorschi S, Ebbets-Reed D, Erickson MR, Yen FT, Bihain BE, Lodish
HF (2001) Proteolytic cleavage product of 30-kDa adipocyte complement-related protein
increases fatty acid oxidation in muscle and causes weight loss in mice. Proc Natl Acad Sci
USA 98:2005–2010
94 J.W. Kuhbier et al.
18. Trayhurn P, Hoggard N, Mercer JG, Rayner DV (1999) Leptin: fundamental aspects. Int J
Obes Relat Metab Disord 23(Suppl 1):22–28
19. Mizuno H, Hyakusoku H (2003) Mesengenic potential and future clinical perspective of
human processed lipoaspirate cells. J Nippon Med Sch 70:300–306
20. Rodriguez AM, Elabd C, Amri EZ, Ailhaud G, Dani C (2005) The human adipose tissue is a
source of multipotent stem cells. Biochimie 87:125–128
21. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK,
Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells.
Mol Biol Cell 13:4279–4295
22. Planat-Benard V, Silvestre JS, Cousin B, Andre M, Nibbelink M, Tamarat R, Clergue M,
Manneville C, Saillan-Barreau C, Duriez M, Tedgui A, Levy B, Penicaud L, Casteilla L
(2004) Plasticity of human adipose lineage cells toward endothelial cells: physiological and
therapeutic perspectives. Circulation 109:656–663
23. Ashjian PH, Elbarbary AS, Edmonds B, DeUgarte D, Zhu M, Zuk PA, Lorenz HP, Benhaim
P, Hedrick MH (2003) In vitro differentiation of human processed lipoaspirate cells into
early neural progenitors. Plast Reconstr Surg 111:1922–1931
24. Fujimura J, Ogawa R, Mizuno H, Fukunaga Y, Suzuki H (2005) Neural differentiation of
adipose-derived stem cells isolated from GFP transgenic mice. Biochem Biophys Res
Commun 333:116–121
25. Safford KM, Hicok KC, Safford SD, Halvorsen YD, Wilkison WO, Gimble JM, Rice HE
(2002) Neurogenic differentiation of murine and human adipose-derived stromal cells.
Biochem Biophys Res Commun 294:371–379
26. Safford KM, Safford SD, Gimble JM, Shetty AK, Rice HE (2004) Characterization of
neuronal/glial differentiation of murine adipose-derived adult stromal cells. Exp Neurol
187:319–328
27. Banas A, Teratani T, Yamamoto Y, Tokuhara M, Takeshita F, Quinn G, Okochi H, Ochiya T
(2007) Adipose tissue-derived mesenchymal stem cells as a source of human hepatocytes.
Hepatology 46:219–228
28. Seo MJ, Suh SY, Bae YC, Jung JS (2005) Differentiation of human adipose stromal cells into
hepatic lineage in vitro and in vivo. Biochem Biophys Res Commun 328:258–264
29. Timper K, Seboek D, Eberhardt M, Linscheid P, Christ-Crain M, Keller U, Muller B,
Zulewski H (2006) Human adipose tissue-derived mesenchymal stem cells differentiate
into insulin, somatostatin, and glucagon expressing cells. Biochem Biophys Res Commun
341:1135–1140
30. Corre J, Barreau C, Cousin B, Chavoin JP, Caton D, Fournial G, Penicaud L, Casteilla L,
Laharrague P (2006) Human subcutaneous adipose cells support complete differentiation but
not self-renewal of hematopoietic progenitors. J Cell Physiol 208:282–288
31. Kilroy GE, Foster SJ, Wu X, Ruiz J, Sherwood S, Heifetz A, Ludlow JW, Stricker DM,
Potiny S, Green P, Halvorsen YD, Cheatham B, Storms RW, Gimble JM (2007) Cytokine
profile of human adipose-derived stem cells: expression of angiogenic, hematopoietic, and
pro-inflammatory factors. J Cell Physiol 212:702–709
32. Rehman J, Traktuev D, Li J, Merfeld-Clauss S, Temm-Grove CJ, Bovenkerk JE, Pell CL,
Johnstone BH, Considine RV, March KL (2004) Secretion of angiogenic and antiapoptotic
factors by human adipose stromal cells. Circulation 109:1292–1298
33. Kitagawa Y, Korobi M, Toriyama K, Kamei Y, Torii S (2006) History of discovery of human
adipose-deived stem cells and their clinical applications. Jpn J Plast Reconstr Surg 49:1097–
1104
34. Fraser JK, Wulur I, Alfonso Z, Hedrick MH (2006) Fat tissue: an underappreciated source of
stem cells for biotechnology. Trends Biotechnol 24:150–154
35. Oedayrajsingh-Varma MJ, van Ham SM, Knippenberg M, Helder MN, Klein-Nulend J,
Schouten TE, Ritt MJ, van Milligen FJ (2006) Adipose tissue-derived mesenchymal stem
cell yield and growth characteristics are affected by the tissue-harvesting procedure.
Cytotherapy 8:166–177
Isolation, Characterization, Differentiation, and Application of Adipose-Derived Stem Cells 95
36. Prunet-Marcassus B, Cousin B, Caton D, Andre M, Penicaud L, Casteilla L (2006) From
heterogeneity to plasticity in adipose tissues: site-specific differences. Exp Cell Res
312:727–736
37. Peptan IA, Hong L, Mao JJ (2006) Comparison of osteogenic potentials of visceral and
subcutaneous adipose-derived cells of rabbits. Plast Reconstr Surg 117:1462–1470
38. Pettersson P, Van R, Karlsson M, Bjorntorp P (1985) Adipocyte precursor cells in obese and
nonobese humans. Metabolism 34:808–812
39. Roncari DA, Lau DC, Kindler S (1981) Exaggerated replication in culture of adipocyte
precursors from massively obese persons. Metabolism 30:425–427
40. Schipper BM, Marra KG, Zhang W, Donnenberg AD, Rubin JP (2008) Regional anatomic
and age effects on cell function of human adipose-derived stem cells. Ann Plast Surg
60:538–544
41. Shahparaki A, Grunder L, Sorisky A (2002) Comparison of human abdominal subcutaneous
versus omental preadipocyte differentiation in primary culture. Metabolism 51:1211–1215
42. Shi YY, Nacamuli RP, Salim A, Longaker MT (2005) The osteogenic potential of adipose-
derived mesenchymal cells is maintained with aging. Plast Reconstr Surg 116:1686–1696
43. Zhu M, Kohan E, Bradley J, Hedrick M, Benhaim P, Zuk P (2009) The effect of age on
osteogenic, adipogenic and proliferative potential of female adipose-derived stem cells.
J Tissue Eng Regen Med 3:290–301
44. van Harmelen V, Rohrig K, Hauner H (2004) Comparison of proliferation and differentiation
capacity of human adipocyte precursor cells from the omental and subcutaneous adipose
tissue depot of obese subjects. Metabolism 53:632–637
45. Hutley LJ, Herington AC, Shurety W, Cheung C, Vesey DA, Cameron DP, Prins JB (2001)
Human adipose tissue endothelial cells promote preadipocyte proliferation. Am J Physiol
Endocrinol Metab 281:E1037–E1044
46. Fang B, Song YP, Li N, Li J, Han Q, Zhao RC (2009) Resolution of refractory chronic
autoimmune thrombocytopenic purpura following mesenchymal stem cell transplantation: a
case report. Transplant Proc 41:1827–1830
47. Khan WS, Adesida AB, Tew SR, Andrew JG, Hardingham TE (2009) The epitope char-
acterisation and the osteogenic differentiation potential of human fat pad-derived stem cells
is maintained with ageing in later life. Injury 40:150–157
48. Weinzierl K, Hemprich A, Frerich B (2006) Bone engineering with adipose tissue derived
stromal cells. J Craniomaxillofac Surg 34:466–471
49. Ferguson RE, Cui X, Fink BF, Vasconez HC, Pu LL (2008) The viability of autologous
fat grafts harvested with the LipiVage system: a comparative study. Ann Plast Surg
60:594–597
50. Jurgens WJ, Oedayrajsingh-Varma MJ, Helder MN, Zandiehdoulabi B, Schouten TE,
Kuik DJ, Ritt MJ, van Milligen FJ (2008) Effect of tissue-harvesting site on yield of stem
cells derived from adipose tissue: implications for cell-based therapies. Cell Tissue Res
332:415–426
51. Tchkonia T, Giorgadze N, Pirtskhalava T, Tchoukalova Y, Karagiannides I, Forse RA,
DePonte M, Stevenson M, Guo W, Han J, Waloga G, Lash TL, Jensen MD, Kirkland JL
(2002) Fat depot origin affects adipogenesis in primary cultured and cloned human pre-
adipocytes. Am J Physiol Regul Integr Comp Physiol 282:R1286–R1296
52. Hauner H, Entenmann G (1991) Regional variation of adipose differentiation in cultured
stromal-vascular cells from the abdominal and femoral adipose tissue of obese women. Int J
Obes 15:121–126
53. Vasconez LG, de la Torre JI (2006) Abdominoplasty. In: Mathes (ed) Plastic Surgery.
Saunders Elsevier, Philadelphia, pp 87–117
54. Pitanguy I (1967) Abdominal lipectomy – an approach to it through an analysis of 300
consecutive cases. Plast Reconstr Surg 30:384–391
55. Pitanguy I (1964) Dermolipectomy of the abdominal wall, thighs, buttocks, and upper
extremity. In: Converse JM (ed) Reconstructive plastic surgery. Saunders, Philadelphia
96 J.W. Kuhbier et al.