Litchfield T M, Mackenzie C D, Varma M G
Department of Allergy and Allied Respiratory Diseases, Guy's Hospital, London, U.K.
Acta Trop. 1991 Feb;48(4):293-304. doi: 10.1016/0001-706x(91)90017-e.
Microfilariae of the bovine parasite Onchocerca lienalis were maintained in vitro using a tick cell line as a feeder layer, and under the conditions provided would develop to the sausage stage. A number of media supported this achievement with CMRL 1066 producing the highest yields, particularly when used in conjunction with either medium 199, HAM's F-12, RPMI 1640 or Mark's M20. The addition of inactivated foetal calf serum (iFCS) suppressed development; on the other hand, the addition of tryptose phosphate broth (TPB) enhanced it. The pH range of the medium preferred by the developing worms was 7.35-7.85, and the most favourable osmolality lay in the range 360 to 390 mosM/kg. Insect-derived hormones did not improve yields of developed larvae nor promote moulting to the second larval stage. The culture conditions described in this work, which favour parasite survival and development, provide further insights into the physiological requirements of filariae as well as guide lines to achieving successful in vitro maintenance of Onchocerca sp. A morphological examination of the developing larvae was carried out at electron microscopical level.
牛寄生虫链尾盘尾丝虫的微丝蚴可利用蜱细胞系作为饲养层在体外培养，在所提供的条件下可发育至腊肠期。多种培养基支持这一成果，其中CMRL 1066产量最高，尤其是与199培养基、HAM's F - 12培养基、RPMI 1640培养基或Mark's M20培养基联合使用时。添加灭活胎牛血清（iFCS）会抑制发育；另一方面，添加胰蛋白胨磷酸盐肉汤（TPB）则会促进发育。发育中的蠕虫偏好的培养基pH范围为7.35 - 7.85，最适宜的渗透压范围为360至390 mosM/kg。昆虫来源的激素既不能提高发育幼虫的产量，也不能促进其蜕皮至第二期幼虫。这项工作中描述的培养条件有利于寄生虫的存活和发育，为丝虫的生理需求提供了进一步的见解，并为成功在体外维持盘尾丝虫属提供了指导方针。在电子显微镜水平对发育中的幼虫进行了形态学检查。
