211blood contaminants appear in the syringe. This method is successful in obtaining
large volumes of ASCs from the SVF and does not negatively affect cellular expan-
sion experiments.
Using previous literature recommendations, we formulated a harvesting tech-
nique, unique to our purposes. We have isolated, expanded, and differentiated ASCs
successfully, harvesting tissue using the Coleman dry needle aspiration technique
with an average of 1,019,129 cells/ml. Based on what is reported in the literature,
we propose the abovementioned harvesting technique to be the preferred technique
for isolation of adipose-derived SVF and ASCs. According to the literature, it is
understood that this technique will be less invasive than resection while still
producing high yields of SVF and ASCs with multipotency, stemness, and genetic
profi ling not infl uenced by local anesthetics.
10.2.4 Factors Infl uencing ASC Isolation and Expansion
The initial method for isolating ASCs from adipose tissue was pioneered in the
1960s. Minced rat fat pads were extensively washed to remove contaminating
hematopoietic cells, incubated with collagenase and centrifuged to obtain a pellet of
SVF containing a heterogeneous population of cells. The selection for plastic adher-
ent fi broblastic like cells from the SVF concluded this isolation process (Rodbell
1966a , b ; Rodbell and Jones 1966 ). Mesenchymal stem cells resident in human
adipose tissue were fi rst described by Zuk and co-workers in 2001. The initial pro-
cedure of mincing human adipose tissue by hand was simplifi ed by the development
of liposuction surgery. Many stem cell laboratories have developed methods to iso-
late and expand MSCs from various tissue sources including adipose tissue.
Although most of these methods share similarities, there are some that differ signifi -
cantly which leads to the following very important unanswered question within the
stem cell research community. If these different tissue sources and methodologies
are used for the preparation of MSCs, are these MSCs suffi ciently similar to allow
Fig. 10.4 The plunger is
withdrawn completely
from the syringe barrel and
the lipoaspirate is decanted
into a sterile bottle
containing PBS
10 Harvesting and Collection of Adipose Tissue for the Isolation...