Yoshino T P, Laursen J R
Department of Pathobiological Sciences, University of Wisconsin, School of Veterinary Medicine, Madison 53706, USA.
J Parasitol. 1995 Oct;81(5):714-22.
In vitro production of Schistosoma mansoni daughter sporocysts (DS) from miracidium-derived mother sporocysts (MS) was achieved by synxenic larval cultivation with cells of the Biomphalaria glabrata embryonic (Bge) cell line. The in vitro growth and viability of MS cocultured with Bge cells or in Bge cell-conditioned medium were significantly extended beyond that of larvae cultured in fresh medium alone. However, complete DS development and emergence from MS were achieved only in the presence of Bge cells. Introduction of either miracidia or previously transformed MS onto Bge cell monolayers resulted in an initial attachment of cultured cells to sporocysts, followed by a gradual encapsulation of larvae by multiple layers of Bge cells. Sporocysts and their encapsulating cells eventually formed large cellular aggregates, within which MS increased 4-fold in size during the first 20 days of cultivation. The timing of in vitro DS development was somewhat variable; however, in general, early embryo formation, i.e., germ cell aggregates with surrounding primitive epithelium, was first detected at 15-20 days of culture, whereas motile, intra-MS daughter stages were seen at 25-30 days and thereafter. Mature, first generation DS, measuring 136 +/- 46 microns long by 22 +/- 6 microns wide, emerged from MS starting at approximately 30-45 days of initial cultivation. Although the basic morphology and size of emergent, in vitro-derived DS were comparable to those propagated in vivo, there was a large reduction in the in vitro reproductive capacity of the MS and a delay in DS culture development.(ABSTRACT TRUNCATED AT 250 WORDS)
通过与光滑双脐螺胚胎(Bge)细胞系的细胞进行共栖幼虫培养,实现了从毛蚴衍生的母孢子蚴(MS)体外生产曼氏血吸虫子孢子蚴(DS)。与Bge细胞共培养或在Bge细胞条件培养基中培养的MS的体外生长和活力显著延长,超过了仅在新鲜培养基中培养的幼虫。然而,只有在Bge细胞存在的情况下,才能实现MS的完全DS发育和出现。将毛蚴或先前转化的MS引入Bge细胞单层,导致培养细胞最初附着在孢子蚴上,随后多层Bge细胞逐渐包裹幼虫。孢子蚴及其包裹细胞最终形成大的细胞聚集体,在培养的前20天内,MS在其中大小增加了4倍。体外DS发育的时间有些变化;然而,一般来说,早期胚胎形成,即带有周围原始上皮的生殖细胞聚集体,在培养15 - 20天时首次检测到,而活动的、MS内的子代阶段在25 - 30天及之后出现。成熟的第一代DS,长136±46微米,宽22±6微米,从最初培养约30 - 45天开始从MS中出现。虽然体外衍生的DS的基本形态和大小与体内繁殖的DS相当,但MS的体外繁殖能力大幅下降,且DS培养发育延迟。(摘要截断于250字)