Your browser doesn't support javascript.
loading
Cryopreservation of Anopheles stephensi embryos.
James, Eric R; Wen, Yingda; Overby, James; Pluchino, Kristen; McTighe, Shane; Matheny, Stephen; Eappen, Abraham; Hoffman, Stephen L; Billingsley, Peter F.
Afiliação
  • James ER; Sanaria Inc, Rockville, MD, USA. ejames@sanaria.com.
  • Wen Y; Sanaria Inc, Rockville, MD, USA.
  • Overby J; Sanaria Inc, Rockville, MD, USA.
  • Pluchino K; Sanaria Inc, Rockville, MD, USA.
  • McTighe S; Sanaria Inc, Rockville, MD, USA.
  • Matheny S; Department of Dermatology, Walter Reed National Military Medical Center, Rockville, MD, USA.
  • Eappen A; Sanaria Inc, Rockville, MD, USA.
  • Hoffman SL; Sanaria Inc, Rockville, MD, USA.
  • Billingsley PF; Sanaria Inc, Rockville, MD, USA.
Sci Rep ; 12(1): 43, 2022 01 07.
Article em En | MEDLINE | ID: mdl-34997079
ABSTRACT
The ability to cryopreserve mosquitoes would revolutionize work on these vectors of major human infectious diseases by conserving stocks, new isolates, lab-bred strains, and transgenic lines that currently require continuous life cycle maintenance. Efforts over several decades to develop a method for cryopreservation have, until now, been fruitless we describe here a method for the cryopreservation of Anopheles stephensi embryos yielding hatch rates of ~ 25%, stable for > 5 years. Hatched larvae developed into fertile, fecund adults and blood-fed females, produced fully viable second generation eggs, that could be infected with Plasmodium falciparum at high intensities. The key components of the cryopreservation method are embryos at 15-30 min post oviposition, two incubation steps in 100% deuterated methanol at - 7 °C and - 14.5 °C, and rapid cooling. Eggs are recovered by rapid warming with concomitant dilution of cryoprotectant. Eggs of genetically modified A. stephensi and of A. gambiae were also successfully cryopreserved. This enabling methodology will allow long-term conservation of mosquitoes as well as acceleration of genetic studies and facilitation of mass storage of anopheline mosquitoes for release programs.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Criopreservação / Embrião não Mamífero / Anopheles Limite: Animals Idioma: En Revista: Sci Rep Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Criopreservação / Embrião não Mamífero / Anopheles Limite: Animals Idioma: En Revista: Sci Rep Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Estados Unidos
...