Key Laboratory of Marine Genetic Resources, State Key Laboratory Breeding Base of Marine Genetic Resources, Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, 361005, China; Fujian Key Laboratory of Marine Genetic Resources, Xiamen, 361005, China.
Key Laboratory of Marine Genetic Resources, State Key Laboratory Breeding Base of Marine Genetic Resources, Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, 361005, China; Fujian Key Laboratory of Marine Genetic Resources, Xiamen, 361005, China.
Fish Shellfish Immunol. 2019 May;88:28-35. doi: 10.1016/j.fsi.2019.02.054. Epub 2019 Feb 28.
Hemocytes are the major immune cells of crustaceans. New hemocyte production is required throughout the life cycle of these animals to maintain a functional immune system. The mechanism of crustacean hematopoiesis has just begun to be understood and new methods are needed for the investigation of this process. Here we report the directed differentiation of granular cells (GCs) from the hematopoietic tissue (HPT) cells of Cherax quadricarinatus in vitro. We started by providing the cultured HPT cells with different additives to induce possible differentiation. We found that crayfish muscle extract greatly promoted the physical status of the cells and induced the formation of refractile cytoplasmic granules. The transcription of marker genes and the production of functional prophenoloxidase further confirmed the formation of mature GCs. In our experiments, young GCs usually started to develop in ∼2 weeks post induction and over 60% of the cells became mature within 3-4 weeks. This is the first time that the fully differentiation of crustacean hemocytes is accomplished in vitro. It provides a powerful tool for in-depth study of crustacean hematopoiesis.
血细胞是甲壳动物的主要免疫细胞。这些动物在整个生命周期中都需要新的血细胞生成来维持其功能性免疫系统。甲壳动物造血的机制才刚刚开始被理解，需要新的方法来研究这个过程。在这里，我们报告了从淡水小龙虾（Cherax quadricarinatus）造血组织（HPT）细胞体外定向分化为颗粒细胞（GCs）的过程。我们首先向培养的 HPT 细胞提供不同的添加剂，以诱导可能的分化。我们发现，螯虾肌肉提取物极大地促进了细胞的生理状态，并诱导形成有折光性的细胞质颗粒。标记基因的转录和功能前酚氧化酶的产生进一步证实了成熟 GCs 的形成。在我们的实验中，年轻的 GCs 通常在诱导后约 2 周开始发育，超过 60%的细胞在 3-4 周内成熟。这是第一次在体外完成甲壳动物血细胞的完全分化。它为深入研究甲壳动物造血提供了有力的工具。
