Cryopreservation of human adipose tissue-derived stem/progenitor cells using the silk protein sericin.
Adipose tissue-derived stem/progenitor cells (ASCs) have attracted attention as a cell source that replaces marrow stromal cells (MSCs); ASCs may thus have applications in both regenerative medicine and cell transplantation. These medical treatments, however, require a high-quality  supply of human ASCs. Therefore, the cryopreservation methods have been improved by changing a component of a cryopreservation medium. Sericin, a protein hydrolysate (with an average molecular weight of 30 kDa) is very rich in serine. The viability and the adipogenic/osteogenic potential of  human ASCs were tested after freezing in a cryopreservation medium containing sericin. After thawing, the viability of the human ASCs frozen in the cryopreservation medium was found to be more than 95%. The proliferation rate of human ASCs frozen in CELLBANKER 2, and DMEM/Ham's F-12 medium  (serum free) + 10% DMSO, 0.1 mol/L maltose, and 1% sericin was higher than that of the cells frozen in the maintenance medium + 10% DMSO. The adipogenic/osteogenic differentiation capabilities of frozen human ASCs were examined by Oil Red O staining/Von Kossa's method. The human ASCs were  frozen using CELLBANKER 2, and DMEM/Ham's F-12 medium (serum free) + 10% DMSO, 0.1 mol/L maltose, and 1% sericin were positive. In conclusion, the cryopreservation medium containing sericin is therefore considered to have a beneficial effect on freezing human ASCs. This serum-free cryopreservation  medium should be widely used in regenerative medicine, cell transplantation, and biological research.
Department of Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate School of Medicine, Higashi-ku, Nagoya, Japan. ymiyamoto@med.nagoya-u.ac.jp
Yoshitaka
Y
Miyamoto
Koichi
K
Oishi
Hiroshi
H
Yukawa
Hirofumi
H
Noguchi
Masahiro
M
Sasaki
Hisashi
H
Iwata
Shuji
S
Hayashi
D000273:Adipose Tissue / Q000166:cytology*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006801:Humans; D047030:Sericins / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects
D016895:Culture Media, Serum-Free; D047030:Sericins; D004121:Dimethyl Sulfoxide