Keisuke Adachi1,2, Yuta Ohno2, Keitaro Satoh2, Akiko Shitara2, Yasunori Muramathu1, Masanori Kashimata3. 1. Department of Oral Maxillofacial Surgery, Asahi University School of Dentistry, Gifu, Japan. 2. Department of Pharmacology, Asahi University School of Dentistry, Gifu, Japan. 3. Department of Pharmacology, Asahi University School of Dentistry, Gifu, Japan matasan@dent.asahi-u.ac.jp.
Abstract
BACKGROUND/AIM: Cryopreservation of cell lines has been widely used in the laboratory; however, cryopreservation of organs is still considered to be difficult. The submandibular gland (SMG) of fetal mice is one of the best-characterized organs. We investigated the conditions for cryopreserving SMG rudiments. MATERIALS AND METHODS: Embryonic day 13 SMG rudiments were cryopreserved with or without a cryoprotectant. They were thawed and incubated in DMEM/F12 medium. Moreover, the influence of EGF stimulation on the signaling cascade after frozen-thawing the rudiments was analyzed by Western blotting. RESULTS: When SMG rudiments were cryopreserved without a cryoprotectant, all cells in the rudiments died. However, the SMG rudiments that had been preserved in a cryoprotectant showed branching morphogenesis. Additionally, the responsiveness of signaling cascades to EGF did not differ between frozen with a cryoprotectant and non-frozen rudiments. CONCLUSION: Cryopreservation might be a useful technology for preserving tissues from small organs, such as fetal SMG rudiments. Copyright
BACKGROUND/AIM: Cryopreservation of cell lines has been widely used in the laboratory; however, cryopreservation of organs is still considered to be difficult. The submandibular gland (SMG) of fetal mice is one of the best-characterized organs. We investigated the conditions for cryopreserving SMG rudiments. MATERIALS AND METHODS: Embryonic day 13 SMG rudiments were cryopreserved with or without a cryoprotectant. They were thawed and incubated in DMEM/F12 medium. Moreover, the influence of EGF stimulation on the signaling cascade after frozen-thawing the rudiments was analyzed by Western blotting. RESULTS: When SMG rudiments were cryopreserved without a cryoprotectant, all cells in the rudiments died. However, the SMG rudiments that had been preserved in a cryoprotectant showed branching morphogenesis. Additionally, the responsiveness of signaling cascades to EGF did not differ between frozen with a cryoprotectant and non-frozen rudiments. CONCLUSION: Cryopreservation might be a useful technology for preserving tissues from small organs, such as fetal SMG rudiments. Copyright
Authors: Melinda Larsen; Matthew P Hoffman; Takayoshi Sakai; Justin C Neibaur; Jonathan M Mitchell; Kenneth M Yamada Journal: Dev Biol Date: 2003-03-01 Impact factor: 3.582