Morphological and functional characterization of isolated stem cells from the midgut of Chilo suppressalis larvae

作者

Zhou, Yancong; Jia, Biantao; Han, Lanzhi; 更多内容

期刊:JOURNAL OF ASIA-PACIFIC ENTOMOLOGY ISSN:1226-8615 , 年:2019 . 卷:22 . 期:3 页码:982-989

语种: English

原文链接:http://dx.doi.org/10.1016/j.aspen.2019.08.001

摘要

The proliferation and differentiation of stem cell populations allow the midgut to grow/regenerate in lepidopteran insect. Basic epithelial regenerative functions can be assessed in vitro by purifying these stem and mature cell populations. Therefore, we isolated and purified stem and mature cells from the midgut of C. suppressalis larvae by density gradient centrifugation and observed the morphologies of these cells. A flow cytometry method was used to monitor C. suppressalis stem cell proliferation and differentiation under different cell culture conditions. We observed high proportions of the stem and differentiating cells in third- and fourth-instar larvae, respectively, indicating that, in larvae, stem cells rapidly proliferate early in development and are strongly differentiated at late stages. Incubation in medium supplemented with fat body extract and ecdysone resulted in a significantly increased proportion of stem cells, not of the differentiating cells, indicating that co-culture with fat body extract and ecdysone stimulates the proliferation of C. suppressalis stem cells. Viability bioassays showed that Cry1Ab displayed significant cytotoxic effects on the midgut cell culture of C. suppressalis. The proportion of differentiating cells was significantly increased after a 48-h exposure to sublethal doses of Cry1Ab toxin, and peaked at the Cry1Ab concentration of 0.3 mu g/ml, demonstrating that epithelial cells with strong regenerative capacity via the differentiation of stem cells. These results improve our understanding of C. suppressalis stem cell biology and illustrate the potential role of the enhanced midgut regeneration induced by stem cell proliferation or differentiation as a reparation mechanism to Bt toxin.

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关键词
Chilo suppressalis; Stem cell; Mature cell; Proliferation and differentiation; Regeneration; Bt toxin

附加关键词:
HELIOTHIS-VIRESCENS; EPITHELIAL-CELLS; MANDUCA-SEXTA; IN-VITRO; DIFFERENTIATION; PROLIFERATION; RESISTANCE; MOTH; LEPIDOPTERA; GROWTH

作者信息

通讯作者:
     Han, LZ , Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China.; Jia, BT , Shanxi Agr Univ, Agr Coll, Taigu 030801, Peoples R China.
作者机构:
     [1] [ Zhou, Yancong; Han, Lanzhi; Peng, Yufa ] Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China.
     [2] [ Zhou, Yancong; Jia, Biantao ] Shanxi Agr Univ, Agr Coll, Taigu 030801, Peoples R China.
作者邮箱:
     jiabtao@foxmail.com
     lzhhan@sina.com