REVIEWMultipotential differentiation of adipose tissue-derived
stem cells
Brian M Strem,1,2 Kevin C Hicok,1 Min Zhu,1 Isabella Wulur,1 Zeni Alfonso,1 Ronda E Schreiber,1John K Fraser1 and Marc H Hedrick1,2
1Cytori Therapeutics, Inc., San Diego, CA, USA
2Departments of Surgery and Bioengineering, University of California, Los Angeles, CA, USA(Received for publication on February 14, 2005)(Revised for publication on May 4, 2005)(Accepted for publication on May 19, 2005)

Abstract. Tissue engineering offers considerable promise in the repair or replacement of diseased
and/or damaged tissues. The cellular component of this regenerative approach will play a key role inbringing these tissue engineered constructs from the laboratory bench to the clinical bedside. However,
the ideal source of cells still remains unclear and may differ depending upon the application. Current
research for many applications is focused on the use of adult stem cells. The properties of adult stem
cells that make them well-suited for regenerative medicine are (1) ease of harvest for autologoustransplantation, (2) high proliferation rates for ex vivo expansion and (3) multilineage differentiation
capacity. This review will highlight the use of adipose tissue as a reservoir of adult stem cells and draw
conclusions based upon comparisons with bone marrow stromal cells. (Keio J Med 54 (3): 132–141,
September 2005)
Key words: adipose tissue-derived stem cells, cell differentiation

Introduction
White adipose tissue has the ability to dynamically
expand and shrink throughout the life of an adult. This
capacity is mediated by the presence of vascular andnon-vascular cells that provide a pool of stem and progenitor cells with unique regenerative capacity. This
review will describe the recent preclinical research focused on these stem cells and identify potential clinicalapplications.
Over the past 25 years bone marrow mesenchymal
stem cells (also frequently referred to as marrow stromal cells and herein referred to as MSCs) have been
the subject of considerable research. These cells have
many properties that suggest considerable potential
utility in cellular therapy for a variety of disorders. For
example, their ability to differentiate into osteoblasts,
both in vitro and in vivo, has led to their clinical use
in pilot studies of an inherited bone disorder, Osteogenesis imperfecta1 while their ability to promoterevascularization following ischemic injury has led topreclinical studies in large and small animal models of

myocardial ischemia.2,3
Like marrow, adipose tissue is a mesodermallyderived organ that contains a stromal population containing microvascular endothelial cells, smooth muscle
cells and stem cells.4 These cells can be enzymaticallydigested out of adipose tissue (commonly from lipoaspirate) and separated from the buoyant adipocytes
by centrifugation. A more homogeneous population
emerges in culture under conditions supportive of
MSC growth. This population (termed Adipose TissueDerived Stem Cells, ADSCs) shares many of thecharacteristics of its counterpart in marrow including
extensive proliferative potential and the ability to undergo multilineage differentiation.5,6
Clonogenic assays have typically been used to quantify MSCs in marrow.7–10 In these assays (colony forming units, CFU-F), cells are plated at @1,000/cm 2 andgrown for two-three weeks. Colonies of more than 50
cells are then quantiﬁed. Using these assays, the number of MSCs in bone marrow is generally found to beapproximately 1 in 25,000 to 1 in 100,000,7–10 althoughmany authors have found this frequency is inﬂuenced

Reprint requests to: Dr. Brian M. Strem, Cytori Therapeutics, Inc., 3020 Callan Road, San Diego, CA, 92121 USA
132

Keio J Med 2005; 54 (3): 132–141

by factors such as age, gender, presence of osteoporosis, and prior exposure to high dose chemotherapy orradiation.7,11–13 Our unpublished data suggest that the
average frequency of ADSCs in processed lipoaspirate(56 donors, median age 49) is @2% of nucleated cells(manuscript in preparation). As with bone marrow stem
cells, yield is dependent upon donor age, body mass index and tissue harvest site. This ﬁnding is consistent
with previous reports showing rapid growth of ADSCs
from cultures initiated at considerably lower cell density than that typically used with marrow.6,14 Further,our studies have indicated that the yield of ADSCs isapproximately 5,000 CFU-F per gram of adipose tissue.
This compares with estimates of approximately 100–
1,000 CFU-F per milliliter of bone marrow.
The signiﬁcance of this difference for stem cell therapy is clear. Donor site morbidity limits the amount ofmarrow that can be obtained and thereby extends thetime in culture required to generate a therapeutic celldose. Thus, the volume of human marrow taken underlocal anesthesia is generally limited to no more than 40
ml and yields approximately 1:2  10 9 nucleated cells.15
Obtaining a larger volume necessitates the use of general anesthesia, increases donor site morbidity,16,17 andfurther dilutes the stem cell fraction with stem cell-freeblood.15 At the stem cell frequency cited above,7,8 this
40 ml of marrow will contain approximately 2:4  10 4
MSCs in a skeletally mature adult. By contrast, a typicalharvest of adipose tissue, under local anesthesia, caneasily exceed 200 ml and yield @2  10 8 nucleated cells
per 100 ml of lipoaspirate.18 Thus, 200 ml lipoaspirate
will typically yield in excess of 1  10 6 stem cells; a differential of approximately 40-fold more than that present in 40 ml of marrow.
Cell Surface Characterization
The cell surface phenotype of human ADSCs is quite
similar to MSCs (see Table).5,19 It is worth notingsome of the key similarities and differences between
these cells. For instance, CD105, STRO-1 and CD166(ALCAM) are three common markers used to identify
cells with multilineage differentiation potential and areconsistently expressed on ADSCs and MSCs.20–26 Also,
CD117 (the stem cell factor receptor) has been shown
to be expressed on an array of totipotent or pluripotent
cells including embryonic stem cells, hematopoietic
stem cells, MSCs and ADSCs.27–29 In addition to these
multipotent markers, ADSCs and MSCs display numerous other molecules including CD29 (beta-1 integrin, which plays a critical role in therapeutic angiogenesis30), CD44 (hyaluronate receptor, which is
crucial in the development of neoextracellular matrix
and plays a role in numerous pathologic and physiologic events) and CD49e (alpha-5 integrin, important

133
Table The Cell Surface Phenotype of ADSCs is Similar
to That of Bone Marrow MSCs
Surface marker

CD9
CD10
CD13
CD29
CD31
CD34
CD44
CD45
CD49d
CD49e
CD54
CD55
CD59
CD90
CD105
CD106
CD117
CD146
CD166
STRO-1

Expression pattern
ADSCs

MSCs

þ
þ
þ
þ


þ

þ
þ
þ
þ
þ
þ
þ

þ
þ
þ
þ

þ
þ
þ
þ


þ


þ
þ
þ
þ
þ
þ
þ
þ
þ
þ
þ

for cell adhesion to ﬁbronectin). ADSCs also express
high levels of CD54 (ICAM-1) when compared with
BM-MSCs.19 ICAM-1 is a member of the immunoglobulin supergene family and can be up-regulated in
response to numerous inﬂammatory mediators andcytokines.31
ADSCs lack the expression of known hematopoietic
and endothelial markers such as CD3, CD4, CD11c,
CD14, CD15, CD16, CD19, CD31, CD33, CD38, CD56,
CD62p, CD104, and CD144. Also, less than 1% of
ADSCs express the HLA-DR protein and the majority
express MHC Class I molecules,18 suggesting their potential for allogeneic transplantation.32 One differencein surface marker expression appears to be the reciprocal expression of VLA-4 (CD49d/CD29) and its
cognate receptor VCAM-1 (CD106). Thus, we haveobserved expression of VLA-4 but not VCAM-1 by
ADSCs from the majority of donors. This is the reverseof the expression pattern of these molecules by
MSCs.19 While the nature of this difference needs further investigation, this observation is intriguing sincethese molecules are involved in hematopoietic stem andprogenitor cell homing to and mobilization from thebone marrow.33,34
Multilineage Differentiation Capacity
We and others have demonstrated the ability of adipose tissue-derived stem cells to undergo differentiation

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Strem BM, et al: Adipose tissue-derived stem cells

FigureAdipose Tissue-Derived Stem Cells are multipotent, extending beyond the traditional mesenchymal lineages.

along classical mesenchymal lineages: adipogenesis,chondrogenesis, osteogenesis, and myogenesis.4–6,35–43
Non-mesenchymal lineages have also been investigatedand conﬁrm the transdifferentiation ability of
ADSCs44–47 (see Figure). Our data using single cellderived clonal populations demonstrates that at least
part of this plasticity resides in a population of multipotential cells.5 In most respects, these data demonstrate a set of functional properties that is very similar,
though not always identical, to that of MSC.
Mesenchymal lineages
Adipogenesis
Given the origin of ADSCs it is not surprising that,when cultured in adipogenic medium, ADSCs express
several adipocytic genes including lipoprotein lipase,aP2, PPAR(gamma)2, leptin, Glut4, and develop lipidladen intracellular vacuoles, the deﬁnitive marker ofadipogenesis.4,5,40,48 Despite certain donor-to-donor
qualitative differences in adipogenic potential,48 thepattern of expression of these genes appears to be very
similar, if not identical, to that observed for adipogenic
differentiation in MSCs.23,49,50
The in vivo capacity of these cells to differentiate
into cells of the adipocytic lineage has also been
demonstrated in studies involving implantation of cellseeded natural (collagen and hyaluronic)51–53 or syn-thetic bioresorbable (polylactic acid, polyglycolicacid)54,55 scaffolds. It is important to note that thesestudies generally agree that robust ectopic in vivo
adipogenesis requires in vitro pre-differentiation of
ADSCs prior to implantation. This requirement may beeliminated by co-implantation of cells with a source ofadipogenic stimuli. Thus, Yuksel and colleagues havedemonstrated ectopic adipogenesis at the site of implantation of microbeads containing insulin and insulinlike growth factor 1.56 This suggests the recruitment of
adipocytic stem and progenitor cells to the site of implantation, however it is not clear if this is a local
recruitment or derives from distal compartments ofsuch progenitors in fat and/or marrow.
Osteogenesis
The ability of MSCs to give rise to osteoblasts is wellknown.57–60 Observations of patients with a rare disorder, progressive osseous heteroplasia, in which calciﬁed nodules form in subcutaneous adipose depots,
provided physiological evidence that similar cells may
exist in adipose tissue.61,62 In the past decade, several groups have isolated cells from adipose tissue ofhumans and other species capable of differentiating into
osteoblasts in vitro.5,36,39,42,63–67 Under osteogenic
conditions, similar to those used for MSCs, ADSCs areobserved to express genes and proteins associated withan osteoblasts phenotype, including alkaline phospha-Keio J Med 2005; 54 (3): 132–141

tase, type I collagen, osteopontin, osteonectin, osteocalcin, bone sialo protein, RunX-1, BMP-2, BMP-4
BMP receptors I and II, PTH-receptor.5,39 ADSCs arealso able to form mineralized matrix in vitro in both
long term 2-D or 3-D osteogenic cultures.
Using a variety of supportive scaffolds, human
ADSCs can form bone in immunodeﬁcient rodent
ectopic bone models.35,55,68 Histologic analysis of these
implants revealed cells staining positive for the humannuclear antigen, conﬁrming that the cells making the
new bone are donor derived. ADSC-derived were
found lining the regions of ectopic bone and also
embedded within the newly formed bone. In a murine
critical size calvarial defect model, ADSCs were able toregenerate cranial bone, which spanned the defect site,
within 8 to 12 weeks of implantation.14 Bone formation
occurred via intramembranous ossiﬁcation, conﬁrmed
by a lack of chondrogenic matrix. Since the calvarium
normally develops through this mechanism, ADSCs
differentiated appropriately for cranial membranous
bone repair. Overall, the rate and extent of bone formation was comparable to that of MSCs.
These ﬁndings supported a recent clinical report
describing successful treatment of a large, bilateralcalvarial defect in a seven year old girl. Repair wasachieved through an approach in which autologous
adipose-derived cells, iliac crest bone, and ﬁbrin glue
were combined with a resorbable mesh.69 CT scanstaken 3 months after surgery showed marked ossiﬁcation throughout the defect. Further, the child no longer
has to wear a protective helmet. This anecdotal reportsuggests that use of adipose-derived cells was safe andthat they may have contributed to healing of a large,
previously refractory defect.
Chondrogenesis
High density micromass cultures of ADSCs andMSCs generate cellular nodules that produce large
amounts of cartilage-related extracellular matrix molecules including sulfated proteoglycans, collagen II and
IV, PRELP and aggrecan.4,5,23,36,38,40,41,70 ADSCs
seeded onto alginate discs and implanted into immunodeﬁcient mice exhibit prolonged (12 weeks) synthesis of
cartilage matrix molecules including collagen II, collagen VI, and aggrecan.38 The identiﬁcation of whichcells (ADSCs or MSCs) are better for engineering cartilage is still under debate. We have analyzed ADSCs
and MSCs from the same patients under identical conditions and found that ADSCs had greater chondrogenic ability than MSCs.6 However, Winter et al. demonstrated that, while ADSCs and MSCs are notsigniﬁcantly different in their ability to undergo chondrogenesis in two-dimensional cultures, MSCs exhibitedenhanced chondrogenesis in three dimensional cul-

135

tures.41 This is in contrast to a recent in vivo study ofmature cartilage development from ADSCs in a rabbit
osteochondral defect model.43 In this study the authors
noted that the repair induced by ADSCs was superior
to that derived from osteochondral autografts. Speciﬁcally, the Pineda score, which assesses four different
parameters of cartilage repair, was greater for ADSCderived grafts than for osteochondral grafts at eachtime point examined. ADSC grafts also showed superior performance in creep indentation biomechanicaltesting performed at 24 weeks. However, it should be
noted that performance at 24 weeks was still inferior to
intact cartilage. Nonetheless, these data indicate thatthe deﬁcit observed by Winter et al. may be a culture
artifact rather than an inherent limitation in the osteochondral capacity of ADSCs.
Non-mesenchymal lineages
Skeletal myogenesis
Culture of ADSCs and MSCs in the presence of
dexamethasone, hydrocortisone, and/or 5 azacytidine
results in a time-dependent pattern of expression ofmuscle-related genes that is consistent with normal
myogenesis. This is deﬁned by early expression of key
master regulatory factors MyoD1 and myf5, myf6, andmyogenin followed by later expression of myosin heavy
chain.5,37,71 This process is associated with characteristic changes in cell morphology including generation oflong, multinucleate, MyoD1-positive cells early in culture and bundles of myosin heavy chain-positive myoﬁbrillar structures appearing after two weeks.
Thus far, only one study has investigated the useof ADSCs to restore functional capacity of damaged
skeletal muscle in vivo. Bacou and colleagues demonstrated that ADSCs transplanted into injured regions ofrabbit skeletal muscle increased muscle weight and ﬁber cross section area, and signiﬁcantly raised the maximal contractile force compared with damaged control
muscles.72 The transplanted cells were identiﬁed in theregion of damaged tissue and some expressed the skeletal muscle markers merosin, myogenin and multiple
isoforms of myosin heavy chain, suggestive of myogenic
differentiation.
Cardiac
Cardiovascular disease (CVD) has emerged as aleading cause of morbidity and mortality, accounting
for approximately one-third of all deaths in 2002.73
Current therapies for CVD are focused on revascularization, followed by drug therapy to mitigate the deleterious remodeling that ensues post ischemia. While
this therapeutic regimen has had a substantial impact

136

on how CVD is treated, there is still a necessity tofurther advance treatment options. Cell therapy for
cardiovascular diseases was originally thought to focus
on the regeneration of injured myocardium. The useof progenitor/stem cells has changed this paradigm
by potentially contributing to multiple mechanisms for
cardiac repair. These mechanisms include, but are notlimited to: increasing blood ﬂow to the ischemic tissue(either by differentiating into neovasculature or secreting factors to recruit or expand endogenous vasculature), reducing cardiac myocyte apoptosis through aparacrine-mediated response, regulating the inﬂammatory milieu (which can alter the negative remodelingthe ensues following acute ischemic injury), or byrecruiting endogenous stem cells (i.e. from the heart74
or bone marrow75) to assist in regenerating the injured
tissue.
While all of these paths could potentially augment
current therapies, regeneration of lost myocardium isstill the ideal outcome, particularly in congestive
heart failure, which involves currently irreversible
loss of contractile function. Both ADSCs45,76,77 andMSCs78–81 have been shown to be capable of in vitro
differentiation into cardiac myocytes. The most compelling data was obtained by Planat-Bernard et al. in
which fresh adipose-derived cells were plated into
semisolid culture. After three weeks, colonies of spontaneously beating cells were observed. These cells
exhibited several molecular, electrophysiologic, andpharmacologic properties of cardiac myocytes.76 However, there is a lack of data addressing the ability of
ADSCs to regenerate damaged myocardium in vivo. In
work presented at the European Society of Cardiology
in Munich, Germany in 2004, Valina et al. presented
data showing that delivery of either ADSCs or MSCs
through the left coronary artery following an acute
myocardial infarction in pigs resulted in improved left
ventricular function after 4 weeks compared to salinetreated controls. Upon histologic analysis, the group
identiﬁed donor ADSCs at the site of infarction, andthat these cells expressed cardiomyocytic markers.
Neurogenesis
In vitro differentiation along the neuronal lineages
has also been demonstrated for both ADSCs5,46,82 andMSCs.83–86 Thus, treatment of rat and human MSCs or
ADSCs with beta-mercaptoethanol results in rapidtransition of cells to a neuronal morphology (a condensed cell body with multiple neuron-like outgrowths), and expression of neural markers including
nestin, neuron-speciﬁc enolase (NSE), and neuronspeciﬁc protein (NeuN), all of which are early markersof the neuronal lineage. Similar results are seen withalternate inductive conditions such as isobutylmethy-

Strem BM, et al: Adipose tissue-derived stem cells

lxanthine (IBMX) and dibutyryl cAMP or forskolin andbutylated hydroxyanisole. We have also detected expression of trkA (a receptor for NGF) and the presenceof voltage gated potassium channels. However, to date,detection of neural markers in in vitro differentiated
ADSCs and MSCs has been restricted to these earlygenes; no expression of markers characteristic of mature neurons, oligodendrocytes, or astrocytes has been
described. This may suggest that the expression of thesemarkers is the result of disordered gene expression
resulting from the toxic inductive stimulus or that theinduction is unmasking an inherent neuronal potentialthat is only partially supported by the culture conditions.
The latter interpretation is supported by in vivo
studies in which ADSCs and MSCs have been
implanted into the CNS of experimental animals. Zhao
et al.87 have demonstrated that implantation of MSCs
into the cortex of rats following ischemic injury resulted
in signiﬁcantly improved performance in a limb placement test and that the implanted cells had undergone
an in vivo change in marker expression consistent withdifferentiation along astrocytic, oligodendrocytic, andneural lineages. However, there was no evidence forincorporation of these cells into the cerebral architecture. Therefore, it is possible that the observed functional improvement was due to an indirect mechanism,for example, paracrine expression of angiogenic and/or
anti-apoptotic factors by the implanted cells would
promote survival of functionally compromised but
viable host tissue.88,89 Other studies have also demonstrated engraftment of MSCs into the region in theabsence of injury albeit with modest evidence of differentiation.90,91 Nonetheless, these data provide substantial support for a potential role of MSCs in direct orindirect (gene-modiﬁed) therapy for the CNS.92,93
ADSCs have also been applied in the setting of experimental stroke. Thus, Kang et al. directly implanted
ADSCs into the brain of rats following 90 minutes of
middle cerebral artery occlusion.94 In some studies, the
ADSCs were transduced with either the lacZ gene or
the gene encoding brain-derived neurotrophic factor(BDNF) as both a marker and a potentially therapeutic
agent. Marked cells were seen throughout the infarct
area 14 days after implantation and a fraction of thesecells co-expressed MAP2 (4% of marked cells) or
GFAP (9% of marked cells) suggesting a degree ofneural differentiation. No data were presented with regard to potential fusion between donor and recipient
cells. As with the MSC studies, ADSC-treated animals
showed signiﬁcant improvement in neurologic testingwith the animals receiving BDNF-transduced cells
exhibiting signiﬁcantly better recovery of function thanthose treated with unmodiﬁed ADSC. The same groupalso examined the co-culture of human ADSCs andmurine NSCs, and observed that when NSCs were cul-

Keio J Med 2005; 54 (3): 132–141

tured on mitomycin-treated ADSC-monolayers, their
proliferation was decreased, but neural differentiation
was increased. However, when NSCs were cultured
with ADSC-conditioned medium or co-cultured withpermeable ﬁlter on which ADSCs were grown, theproliferation of NSCs signiﬁcantly decreased and glial
differentiation increased, but their neural differentiation was not affected. Thus Kang et al. concluded that
direct physical contact between ADSCs and NSCs isrequired for induction of neural differentiation.95
Other applications
Therapeutic angiogenesis
Restoring blood ﬂow to ischemic tissue has proveninstrumental in the treatment of patients with acute
myocardial infarctions. However, this application is not
the only one that could greatly beneﬁt by increased
angiogenesis. Therapeutic angiogenesis, potentiallyfrom ADSCs and MSCs, has an extremely broad rangeof clinical applications under investigation, such as
ischemic cardiomyopathy, peripheral vascular disease,ischemic stroke, acute tubular necrosis, diabetic retinopathy, ischemic encephalopathy, traumatic spinal cord
injuries, and transplant related ischemia. Both ADSCs
and MSCs have been shown to increase angiogenesis toischemic tissue; however the underlying mechanism
remains unclear. Both cell types have been shown to
excrete substantial quantities of angiogenic growth
factors, including VEGF, PlGF, bFGF, angiogenin,
GM-CSF, MCP-1 and SDF-1alpha.96,97 Both cell types
have also been shown to be capable of endothelial
differentiation.44,98
Miranville et al. have presented data supporting thepresence of cells within adipose tissue that differentiated into endothelium.99 Thus, CD34þ /CD31 cells
within adipose were shown to be capable of in vitro
differentiation into cells that expressed both CD31 andvon Willebrand factor, both markers of mature endothelium. Most importantly, the authors demonstrated
the ability of these cells to improve blood ﬂow andcapillary density in a NOD-SCID mouse model of hindlimb ischemia. These data are conﬁrmed by anotherstudy showing that delivery of ADSCs to immunodeﬁcient animals following induction of severe hind limb
ischemia results in accelerated restoration of perfusion.96 As an interesting side note, Miranville et al.
also reported that approximately 18% of CD34þ /CD31 cells co-expressed ABCG2, a protein associated with the side population (SP) stem cell phenotype; overall approximately 4% of all non-buoyant
adipose-tissue-derived cells express ABCG2.99 It
should be noted that there, at present, are no data
addressing the question of whether or not the endothe-137

lial and mesenchymal differentiation capacity within
adipose tissue reside within the same cells.
Gene delivery
A number of investigators have transduced ADSCs
in order to facilitate tracking or to engender a therapeutic effect. Thus, Leo et al. used Ad-CMV-luciferase
to allow non-invasive, real-time tracking of ADSCs in
rat spine.100 Similarly, Dragoo et al.35 infected both
MSCs and ADSCs with E1A-deleted-type 5 adenovirus
constructs containing the BMP-2 (bone morphogenic
protein-2) gene or the bacterial beta-galactosidase(lacZ) gene. LacZ gene transduction efﬁciency was
35% for MSCs and 55% for ADSCs. Ad-BMP2 infection resulted in levels of expression of BMP-2 proteinthat were three-fold higher than those derived from
MSCs. Ad-BMP-2 infected ADSCs exhibited in vitro
osteoblastic differentiation in the absence of exogenous
osteogenic factors. They also exhibited robust ectopic
in vivo production of bone when cells were implanted
into a collagen sponge within the subcutaneous space.35
Given the success of unmodiﬁed MSCs in treatment of
osteogenesis imperfecta101,102 these data support thepotential for transplant of allogeneic or gene-modiﬁed
ADSCs for genetic disorders of the skeletal system.
Kang et al.94 have also used an E1A-deleted type 5
adenovirus to infect ADSCs. As described above, thesestudies employed transduction of a tracking gene (lacZ)and a potentially therapeutic gene Brain-Derived Neurotrophic Factor (BDNF) achieving 100% and 94%
transduction efﬁciency, respectively. Transduced cells
were implanted into areas of the brain that had undergone transient (90 minute) ischemia-reperfusion injury.
Donor cells capable of continued expression of thetransgene were retained to 30 days, the longest time
point examined in this study.
Finally, our group has published results of a study
comparing infection of ADSCs with oncoretroviral, andlentiviral vectors.103 The primary lentivirus used was
the VSV-G pseudotyped HIV-1 vector SIN18-RhMLV-E (VSV). Infection by the VSV-G pseudotyped
MuLV oncoretrovirus SR(alpha)L-EGFP and a second
lentivirus construct, RRL-PGK-EGFP-SIN18, was also
used. Direct comparison of infection of the lentiviruses
and the oncoretrovirus was possible due to common
envelope protein and the similarity of the transcription
level driven in transduced cells. Thus, we were able toinfect ADSCs with the same number of EGFP transduction units (virus preparations standardized to drive
the same level of GFP expression in control cells) anddetermine efﬁciency by ﬂow cytometry three days andone week after infection. The lentiviral contructs
resulted in 4–10-fold higher expression than the retroviral vector. The percentage of transduced cells was

138Strem BM, et al: Adipose tissue-derived stem cells

not high (10–15%) but remained stable in culture over
100 days. Moreover, using a lentiviral vector with thecytomegalovirus promoter resulted in a transduction
efﬁciency of >90% at a MOI of 14. Studies using
lentiviral-infected cells (RRL-PGK-EGFP-SIN18; MOI59) in which transduction efﬁciency was 98% at day 3
and >95% at day 100 allowed examination of gene
expression during in vitro differentiation. Retention ofmarker gene (EGFP) expression was observed following both adipogenic and osteogenic differentiation.
Hematopoietic support
The ability to support hematopoiesis is anotherproperty of MSCs that may be important in clinicalapplications. Indeed, co-infusion of MSCs with grafts
containing hematopoietic stem cells has been shown toenhance the rate of hematopoietic engraftment in human clinical studies.104 Hematopoietic support is also
important in transduction of CD34-positive hematopoietic stem and progenitor cells.105–107 While nostudy to date has speciﬁcally examined the ability of
ADSCs to support hematopoiesis, a recent study has
claimed to demonstrate that adipose tissue contains a
population of cells with hematopoietic stem cell activity; that is, a population of cells capable of rescuing
lethally irradiated animals.108 Thus, intraperitonealtransplant of 10 7 fresh adipose or marrow cells was
associated with 40% survival following 10 Gy irradiation. Recovery of platelet and white blood cells counts
was more rapid with marrow than for adipose tissue
cells (8 weeks vs 10 weeks for return to normal levels).
However, adipose-derived cell transplant resulted in avery low level of hematopoietic chimerism (1.7% marrow hematopoietic progenitor cells of donor origin).
This suggests that the survival advantage conferred byadipose-derived cell transplant is due to enhancementof the recovery of endogenous hematopoietic stem cells
from the otherwise lethal irradiation in a manner that is
generally consistent with the human co-infusion study
cited above.104
Summary
In summary, adipose tissue, like bone marrow,contains a population of cells that has extensive selfrenewal capacity and the ability to differentiate along
multiple lineages. The cells possessing this activity canbe obtained in large numbers at high frequency from atissue source that can be extracted in large quantities
with minimal morbidity, unlike marrow. These cells canalso be infected by adenoviral, oncoretroviral, and lentiviral vectors with moderate to high efﬁciency. Thus,
adipose tissue appears to represent a potential clinicallyuseful source of cells for cellular therapy, tissue engineering and gene transfer applications.
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