Cryopreservation of mouse embryoid bodies.
Cryopreservation of embryonic stem (ES) cells is essential to establish them as a resource for regenerative therapy. We evaluated survival adhesion rate, cell structure, gene expression, and multipotency of frozen and thawed embryoid bodies (EBs) derived from mouse ES cells. EBs were cryopreserved using the BICELL and the Program Freezer. After one week the EBs were thawed and cultured. EBs prepared in the Program Freezer had the highest survival adhesion (Program Freezer; 55-69%, BICELL; 30-38%). Though many cells in the thawed EBs were damaged, some were not, especially those prepared in the Program Freezer. RT-PCR analysis showed that genes characteristic of the three embryonic germ layers were expressed in thawed EBs cultured for one week. EBs transplanted into mice formed teratomas consisting of cells derived from the three germ layers. In conclusion, EBs frozen in the Program Freezer had higher survival adhesion rates compared to the BICELL and formed differentiated cells characteristic of the three embryonic germ layers.
Ichikawa
Hinako
H
Department of Anatomy and Organ Technology, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan. hinako-i@sch.md.shinshu-u.ac.jp
No
Heesung
H
Takei
Shunsuke
S
Takashimizu
Itsukei
I
Yue
Fengming
F
Cui
Li
L
Ogiwara
Naoko
N
Johkura
Kohei
K
Nishimoto
Yoshiteru
Y
Sasaki
Katsunori
K
D000818:Animals; D002454:Cell Differentiation; D015925:Cryopreservation / Q000379:methods*; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000502:physiology*; D005855:Germ Layers / Q000648:ultrastructure; D051379:Mice