无血清培养基中 的连续体外培养。
Continuous In Vitro Culture of in a Serum-Free Medium.
机构信息
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
出版信息
Cells. 2023 Feb 2;12(3):482. doi: 10.3390/cells12030482.
Human babesiosis is an emerging tick-borne disease, caused by haemoprotozoa genus of . Cases of transfusion-transmitted and naturally acquired infection have been reported worldwide in recent years and causing a serious public health problem. is one of the important pathogens of human babesiosis, which seriously endangers human health. The in vitro culture systems of have been previously established, and it requires fetal bovine serum (FBS) to support long-term proliferation. However, there are no studies on serum-free in vitro culture of . In this study, we reported that achieved long-term serum-free culture in VP-SFM AGT (VP-SFM) supplemented with AlbuMax I. The effect of adding different dilutions of AlbuMax I to VP-SFM showed that 2 mg/mL AlbuMax I had the best growth curve with a maximum percentage of parasitized erythrocytes (PPE) of over 40%, and it can be used for long-term in vitro culture of . However, the commonly used 20% serum-supplemented medium only achieves 20% PPE. Clearly, VP-SFM with 2 mg/mL AlbuMax I (VP-SFMA) is more suitable for the in vitro proliferation of VP-SFM supplemented with CD lipid mixture was also tested, and the results showed it could support the parasite growth at 1:100 dilution with the highest PPE of 40%, which is similar to that of 2 mg/mL AlbuMax I. However, the CD lipid mixture was only able to support the in vitro culture of for 8 generations, while VP-SFMA could be used for long-term culture. To test the pathogenicity, the VP-SFMA cultured was also subjected to hamster infection. Results showed that the hamster developed dyspnea and chills on day 7 with 30% PPE before treatment, which is similar to the symptoms with un-cultured . This study develops a unique and reliable basis for further understanding of the physiological mechanisms, growth characteristics, and pathogenesis of babesiosis, and provides good laboratory material for the development of drugs or vaccines for human babesiosis and possibly other parasitic diseases.
人类巴贝斯虫病是一种新兴的蜱传疾病，由血原虫属引起。近年来，全球有报道称存在输血传播和自然感染的病例，这给公共卫生带来了严重威胁。是人类巴贝斯虫病的重要病原体之一，严重危害人类健康。已经建立了的体外培养系统，需要胎牛血清（FBS）来支持长期增殖。然而，目前还没有关于无血清体外培养的研究。在这项研究中，我们报告说在添加了 AlbuMax I 的 VP-SFM AGT（VP-SFM）中实现了的长期无血清培养。添加不同稀释度的 AlbuMax I 对 VP-SFM 的影响表明，2mg/mL 的 AlbuMax I 具有最佳的生长曲线，寄生红细胞百分比（PPE）超过 40%，可用于的长期体外培养。然而，常用的 20%血清补充培养基仅能达到 20%的 PPE。显然，添加 2mg/mL AlbuMax I 的 VP-SFM（VP-SFMA）更适合的体外增殖。VP-SFM 中添加 CD 脂质混合物也进行了测试，结果表明，在 1:100 稀释度下，它可以支持寄生虫生长，最高 PPE 为 40%，与 2mg/mL AlbuMax I 相似。然而，CD 脂质混合物仅能支持培养 8 代，而 VP-SFMA 则可用于长期培养。为了测试致病性，用 VP-SFMA 培养的也进行了仓鼠感染。结果表明，仓鼠在治疗前第 7 天出现呼吸困难和寒战，PPE 为 30%，与未培养的相似。这项研究为进一步了解巴贝斯虫病的生理机制、生长特性和发病机制奠定了独特而可靠的基础，并为人类巴贝斯虫病和可能其他寄生虫病的药物或疫苗开发提供了良好的实验室材料。
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