MalarVx, Inc, 1551 Eastlake Ave N, Suite 100, Seattle, WA, 98102, USA.
University of Wisconsin-Madison, Madison, WI, USA.
Malar J. 2024 Aug 20;23(1):251. doi: 10.1186/s12936-024-05079-7.
Plasmodium falciparum oocysts undergo growth and maturation in a unique setting within the mosquito midgut, firmly situated between the epithelium and the basal lamina. This location exposes them to specific nutrient exchange and metabolic processes while in direct contact with the mosquito haemolymph. The limited availability of in vitro culture systems for growth of the various P. falciparum mosquito stages hampers study of their biology and impedes progress in combatting malaria.
An artificial in vitro environment was established to mimic this distinctive setting, transitioning from a 2D culture system to a 3D model capable of generating fully mature oocysts that give rise to in vitro sporozoites.
A two-dimensional (2D) chamber slide was employed along with an extracellular matrix composed of type IV collagen, entactin, and gamma laminin. This matrix facilitated development of the optimal medium composition for cultivating mature P. falciparum oocysts in vitro. However, the limitations of this 2D culture system in replicating the in vivo oocyst environment prompted a refinement of the approach by optimizing a three-dimensional (3D) alginate matrix culture system. This new system offered improved attachment, structural support, and nutrient exchange for the developing oocysts, leading to their maturation and the generation of sporozoites.
This technique enables the in vitro growth of P. falciparum oocysts and sporozoites.
疟原虫卵囊在蚊子中肠内的一个独特环境中生长和成熟，牢固地位于上皮细胞和基膜之间。这种位置使它们在与蚊子血淋巴直接接触的同时，暴露于特定的营养交换和代谢过程中。缺乏用于各种疟原虫蚊子阶段生长的体外培养系统，阻碍了对其生物学的研究，并阻碍了疟疾防治工作的进展。
建立了一种人工体外环境来模拟这种独特的环境，从二维培养系统过渡到能够生成完全成熟卵囊并产生体外子孢子的三维模型。
使用二维（2D）腔室载玻片和由 IV 型胶原、entactin 和γ层粘连蛋白组成的细胞外基质。该基质促进了最佳培养基成分的开发，用于体外培养成熟的疟原虫卵囊。然而，这种 2D 培养系统在复制体内卵囊环境方面的局限性促使我们通过优化三维（3D）藻酸盐基质培养系统来改进方法。这个新系统为发育中的卵囊提供了更好的附着、结构支撑和营养交换，从而导致它们成熟并产生子孢子。
该技术可实现疟原虫卵囊和子孢子的体外生长。
