Biochemical and Biophysical Research Communications
Isolation and identification of mesenchymal stem cells from human lipoma tissue
Section snippets
Materials and methods
Cell culture media. The unique feature of our cell culture method is the use of a low calcium medium and agents known to affect cellular redox states, as mentioned below. The mesenchymal stem cells were developed from human lipoma tissue using a modified MCDB 153 medium (Keratinocyte-SFM, Gibco-Invitrogen Corporation) supplemented with 2 mM N-acetyl-l-cysteine (NAC; Sigma A8199) and 0.2 mM l-ascorbic acid 2-phosphate (Asc 2P; Sigma A8960). This medium was referred to as K-NAC medium in our
Development of cell culture from lipoma tissue
After one day, the culture of minced lipoma tissue in the modified Eagle’s medium (D-medium, as described in materials and methods) with 10% FBS showed most of the cells or cell aggregates remaining in suspension, including many erythrocytes. After washing to remove these cells, few attached single cells or cell clumps were observed. After a change to K-NAC medium with 5% FBS, these attached cells proliferated actively and became near confluence after one week in 25-cm2 flasks. Morphologically,
Acknowledgment
We are deeply thankful to the help of Miss Samantha Benton in the preparation of this manuscript.
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These authors contributed equally to this work.