Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation.
BACKGROUND
Human adipose-derived stem (stromal) cells are promising as a regenerative therapy tool for defective tissues of mesenchymal lineage, including fat, bone, and cartilage, and blood vessels. In potential future clinical applications, adipose-derived stem cell cryopreservation could be an indispensable fundamental technology, as has occurred in other fields involving cell-based therapies using hematopoietic stem cells and umbilical cord blood cells.


METHODS
The authors examined the proliferative capacity and multipotency of human adipose-derived stem cells isolated from lipoaspirates of 18 patients in total before and after a 6-month cryopreservation following their defined protocol. Proliferative capacity was quantified by measuring doubling time in cell culture, and multipotency was examined with differentiation assays for chondrogenic, osteogenic, and adipogenic lineages. In addition, expression profiles of cell surface markers were determined by flow cytometry and compared between fresh and cryopreserved adipose-derived stem cells.


RESULTS
Cryopreserved adipose-derived stem cells fully retained the potential for differentiation into adipocytes, osteoblasts, and chondrocytes and for proliferative capacity. Flow cytometric analyses revealed that surface marker expression profiles remained constant before and after storage.


CONCLUSIONS
Adipose-derived stem cells can be cryopreserved at least for up to 6 months under the present protocol without any loss of proliferative or differentiation potential. These results ensure the availability of autologous banked adipose-derived stem cells for clinical applications in the future.
Gonda
Koichi
K
Tokyo and Kanagawa, Japan From the Department of Plastic Surgery, University of Tokyo School of Medicine, and the Division of Research and Development, Biomaster, Inc.
Shigeura
Tomokuni
T
Sato
Takahiro
T
Matsumoto
Daisuke
D
Suga
Hirotaka
H
Inoue
Keita
K
Aoi
Noriyuki
N
Kato
Harunosuke
H
Sato
Katsujiro
K
Murase
Syoko
S
Koshima
Isao
I
Yoshimura
Kotaro
K
D017667:Adipocytes / Q000166:cytology*; D000328:Adult; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D020219:Chondrogenesis; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D005500:Follow-Up Studies; D006801:Humans; D008875:Middle Aged; D010006:Osteoblasts / Q000166:cytology; D010012:Osteogenesis; D010784:Photomicrography; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D014182:Transplantation, Autologous