Morphological and functional characterization of isolated stem cells from the midgut of Chilo suppressalis larvae |
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| Institution: | 1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2. Agricultural College of Shanxi Agricultural University, Taigu 030801, China;1. Department of vegetable crops, Korea National College of Agriculture and Fisheries, Jeonju-si, Jeollabuk-do 54874, Republic of Korea;2. Cheorwon plasma research institute, Cheorwon-gun, Gangwon-do 24047, Republic of Korea;1. Hongcheon Institute of Medicinal Herb, Hongcheon, Gangwon, 25142, Republic of Korea;2. Department of Agricultural Biology, National Institute of Agricultural Science, RDA, Wanju 55365, Republic of Korea;1. Department of Natural and Life Sciences, Faculty of Exact Sciences and Natural and Life Sciences, Larbi Tebessi University, 12002, Tebessa, Algeria;2. Laboratory of Natural Resources and Management of Sensitive Environments ‘RNAMS’, University of Oum-El-Bouaghi, 04000, Oum-El-Bouaghi, Algeria;1. Department of Botany, University of Rajasthan, J. L. N. Marg, Jaipur 302004, Rajasthan, India;2. National Centre for Cell Science, University of Pune Campus, Ganeshkhind, Pune, Maharashtra 411007, India;3. Central Muga Eri Research and Training Institute, (Central Silk Board) Lahdoigarh, Jorhat 785700, Assam, India |
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| Abstract: | 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 μg/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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