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cell viability assessments that there was no substantial difference in cell phenotype
using wet and dry liposuction techniques (Tommaso et al. 2012 ). Keck and col-
leagues ( 2010 ) however noted that local anesthetics have a marked infl uence on the
quality and quantity of viable preadipocytes and ASCs. After the SVF was cultured
for 24–48 h and nonadherent cells washed off, the ASCs were trypsinized and
exposed for 30 min to different anesthetics before being analyzed by fl ow cytome-
try. It was demonstrated that articaine/epinephrine and lidocaine strongly reduced
ASC viability, while bupivacaine had no effect. These exposed cells were then
induced to differentiate into adipocytes for 12 days, and the expression of adiponec-
tin was measured using quantitative real-time polymerase chain reaction. All the
anesthetics tested, namely, bupivacaine, mepivacaine, ropivacaine, articaine/epi-
nephrine, and lidocaine signifi cantly decreased adiponectin expression and adipo-
genic differentiation capacity compared to saline controls. Interestingly,
anesthetic-exposed ASC cultures, except those exposed to articaine/epinephrine,
showed a similar phenotypic appearance to that of control cultures not exposed to
anesthetics. All local anesthetic-exposed ASCs demonstrated impaired adipocyte
differentiation as determined by adiponectin expression. The ASCs from the artic-
aine-/epinephrine-exposed cultures appeared smaller in size, while a similar per-
centage of cells demonstrated lipid droplet formation (Keck et al. 2010 ).
10.2.3.3 Suggested ASC Harvesting Technique
The Coleman technique is a dry needle aspiration procedure and is commonly used
to obtain virgin adipose tissue. No pharmacological substances, e.g., saline and/or
lignocaine (wetting solution), are injected into the donor area before the procedure.
The aim is to induce minimal trauma to the adipocytes and preadipocytes during
collection compared to other harvesting techniques using high negative pressure.
Our group has employed the Coleman technique and restricted all adipose tissue
collections to the use of a 10 ml syringe for the purpose of creating and maintaining
a low negative pressure system. The negative pressure created by a 10 ml syringe
has previously been measured with an aneroid vacuum meter. It was shown to be
510 mmHg when the plunger was withdrawn to a maximum and gradually decreased
as the syringe was fi lled with harvested lipoaspirate (Novaes et al. 1998 ). General
anesthesia is administered for the respective cosmetic procedures, and antiseptic
cleaning of the donor area with chlorhexidine is performed by the theater nurse
prior to the initiation of surgery. Sterility is maintained throughout the procedure.
A small puncture wound (~10 mm) is made with a no. 15 scalpel blade through the
epidermis in the donor areas, i.e., in the infraumbilical and/or fl ank areas (supralat-
eral pelvic area). A Coleman blunt tip harvesting cannula with two distal openings,
which give the tip a shape reminiscent of a bucket handle (Fig. 10.1 ) attached to a
10 ml Luer Lock syringe (syringe), is used to allow for the collection of strands of
adipose tissue. Dry needle aspiration is performed with a harvesting cannula with
dimensions of 150 mm in length, an outer diameter of 4 mm, and inner diameter of
2.5 mm (Johnson & Johnson, Biron 02-331).
F.A. van Vollenstee et al.