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that have been negatively affecting the. With patient's informed consent and IRB approved by the review board of Kaohsiung Medical University Hospital (KMUH-IRB-960443), the hASCs were isolated from subcutaneous adipose tissue from female donors who underwent transverse rectus abdominis myocutaneous (TRAM) flap for breast reconstruction. The hASCs were isolated by standard protocol with some modifications as follows . Tissues were washed with PBS containing 1% penicillin/streptomycin (P/S) (Gibco-BRL, USA). After removing red blood cells, the human adipose tissue was digested overnight in PBS supplemented with 0.01% (w/v) collagenase type I (Gibco-BRL) at 37 °C. The suspension was centrifuged at 1200 RPM for 10 min, and the pellet was washed several times with PBS. The isolated cells were incubated in K-SFM (Invitrogen, USA) supplemented with 10% fetal bovine serum (FBS, Gibco-BRL) and 1% P/S at 37 °C under 5% CO2. The hASCs were tested for potential multi-lineage differentiation of adipogenesis, osteogenesis, and chondrogenesis as previous report [37, 42, 43, 44].
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