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2015 ; 136
(2
): 189e-199e
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Differential Effects of Processing Time and Duration of Collagenase Digestion on
Human and Murine Fat Grafts
#MMPMID26218393
Seaman SA
; Tannan SC
; Cao Y
; Peirce SM
; Lin KY
Plast Reconstr Surg
2015[Aug]; 136
(2
): 189e-199e
PMID26218393
show ga
BACKGROUND: Autologous fat graft retention is unpredictable, and mechanisms of
optimization are poorly understood. Attempts at improving retention use
collagenase experimentally and clinically to isolate the stromal vascular
fraction to "enhance" fat grafts. However, no standardized duration for
collagenase digestion or time following fat graft harvest has been established.
This study investigates the effect of (1) time after fat graft harvest and (2)
collagenase digestion time on interstitial cell and adipocyte viability in murine
fat and human lipoaspirate. METHODS: Murine fat and human lipoaspirate were
incubated ex vivo after harvest at room temperature for 120 minutes. Additional
groups were incubated with collagenase for increasing 5-minute intervals from 30
to 60 minutes. Samples from each group were stained with BODIPY to quantify
intact adipocytes and the LIVE/DEAD kit to quantify interstitial cell viability.
RESULTS: With increased time after harvest, the number of intact adipocytes in
murine fat and human lipoaspirate remained unchanged. Human interstitial cells
were resistant to the effect of increased time ex vivo, whereas murine
interstitial cells decreased in viability. In both populations, increased
collagenase digestion time significantly decreased the number of viable
adipocytes (murine, p ? 0.001; human, p ? 0.001) and interstitial cells (murine,
p ? 0.001; human, p ? 0.001). CONCLUSIONS: Human and murine adipocytes and human
interstitial cells appear resistant to deleterious effects of increasing time
following harvest. However, murine interstitial cells are sensitive to increased
time and prolonged collagenase digestion. These studies highlight the complex
cellular components of fat grafts and how they respond differentially to time and
collagenase digestion.