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In Vitro Cell Dev Biol Anim. 2017 Jun;53(6):532-537. doi: 10.1007/s11626-017-0140-7. Epub 2017 Feb 22.

Relevant principal factors affecting the reproducibility of insect primary culture.

Author information

1
Nihon BioData Corporation, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa, 213-0012, Japan. norichik@nbiodata.com.
2
Laboratory of Applied Entomology, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8, Saiwai-cho, Tokyo, Fuchu, 183-8501, Japan.

Abstract

The primary culture of insect cells often suffers from problems with poor reproducibility in the quality of the final cell preparations. The cellular composition of the explants (cell number and cell types), surgical methods (surgical duration and surgical isolation), and physiological and genetic differences between donors may be critical factors affecting the reproducibility of culture. However, little is known about where biological variation (interindividual differences between donors) ends and technical variation (variance in replication of culture conditions) begins. In this study, we cultured larval fat bodies from the Japanese rhinoceros beetle, Allomyrina dichotoma, and evaluated, using linear mixed models, the effect of interindividual variation between donors on the reproducibility of the culture. We also performed transcriptome analysis of the hemocyte-like cells mainly seen in the cultures using RNA sequencing and ultrastructural analyses of hemocytes using a transmission electron microscope, revealing that the cultured cells have many characteristics of insect hemocytes.

KEYWORDS:

Coleoptera; Interindividual variation; Primary explant culture; RNA-seq; Transmission electron microscope

PMID:
28229336
PMCID:
PMC5486689
DOI:
10.1007/s11626-017-0140-7
[Indexed for MEDLINE]
Free PMC Article

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