TY - JOUR
T1 - Embryonic stage selection for cryopreservation of the silkworm Bombyx mori and the effects of cryopreservation on embryo tissues
AU - Fukumori, Hisayoshi
AU - Yoshida, Mikiyo
AU - Tanaka, Daisuke
AU - Banno, Yutaka
N1 - Funding Information:
This work was supported by the Ministry of Education, Science, Sports and Culture of Japan , National BioResource Project (NBRP) . The work presented here was supported in part as a Collaborative Research Project organized by the Interuniversity Bio-Backup Project (IBBP) of National Institute for Basic Biology.
Funding Information:
We are sincerely grateful to Dr. Y. Mochida and Dr. Y. Takemura of the Institute of Sericulture for their kind help and advice regarding the methods of egg dechorionation and in vitro culture. The use of silkworm strains in this study was supported by the National BioResource Project (NBRP) of the MEXT , Japan. This work was supported by NIBB Collaborative Research Program ( 19-913 ) to YB. We would like to thank Enago ( www.enago.jp ) for the English language review.
Publisher Copyright:
© 2020
PY - 2020/8
Y1 - 2020/8
N2 - Successful cryopreservation of the important silkworm bioresource, Bombyx mori, is essential. In this study, we aimed for successful cryopreservation using vitrification of silkworm embryos. Furthermore, the embryos were assessed for the most appropriate sampling stage. We found that vitrified embryos developed to the serosa ingestion stage when they were vitrified at embryonic stage 24–25. The most suitable stage for vitrification was around a 5–10 h period when the tracheal fibers were elongating in stage 25. None of the vitrified embryos developed into larvae, although some did develop to the pre-hatching stage. From histological analysis, we found that several small cracks formed on the cuticle covering the hypodermis in the vitrified embryos. Additionally, the midgut epithelium was detached from the midgut wall and mixed with the yolk in the midgut lumen. We speculate that the vitrified embryos died from a rapid loss of body water from the small cracks formed in the cuticle. We also suggest that the vitrified embryos may have resulted in dysfunction of the midgut.
AB - Successful cryopreservation of the important silkworm bioresource, Bombyx mori, is essential. In this study, we aimed for successful cryopreservation using vitrification of silkworm embryos. Furthermore, the embryos were assessed for the most appropriate sampling stage. We found that vitrified embryos developed to the serosa ingestion stage when they were vitrified at embryonic stage 24–25. The most suitable stage for vitrification was around a 5–10 h period when the tracheal fibers were elongating in stage 25. None of the vitrified embryos developed into larvae, although some did develop to the pre-hatching stage. From histological analysis, we found that several small cracks formed on the cuticle covering the hypodermis in the vitrified embryos. Additionally, the midgut epithelium was detached from the midgut wall and mixed with the yolk in the midgut lumen. We speculate that the vitrified embryos died from a rapid loss of body water from the small cracks formed in the cuticle. We also suggest that the vitrified embryos may have resulted in dysfunction of the midgut.
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U2 - 10.1016/j.cryobiol.2020.06.016
DO - 10.1016/j.cryobiol.2020.06.016
M3 - Article
C2 - 32621808
AN - SCOPUS:85088086936
VL - 95
SP - 9
EP - 14
JO - Cryobiology
JF - Cryobiology
SN - 0011-2240
ER -