Wisnewski N, Weinstein P P
Department of Biological Sciences, University of Notre Dame, Indiana 46556.
J Parasitol. 1993 Jun;79(3):390-8.
In vitro culture methods were utilized to investigate specific nutritional requirements of Brugia pahangi larvae. Infective third-stage larvae (L3) isolated from Aedes aegypti (Liverpool) mosquitoes were cultured in NCTC 135:IMDM (NI) medium plus various types of serum under 5% CO2/air and 5% CO2/N2 atmospheres. Larvae grew, developed, and molted to the fourth stage (L4); however, further growth and differentiation beyond the mid-L4 were minimal. Serum supplementation was necessary to induce molting of B. pahangi L3s. Iron-supplemented calf serum (FeCS) consistently promoted higher percentages of molting than the other 4 types of serum tested. Specific nutritional requirements for B. pahangi larvae were assessed through testing of supplements known to be important for growth of mammalian cells or other parasites in vitro. Addition of various concentrations and combinations of hemoglobin, insulin, transferrin, selenium, albumin, cholesterol, or a cell feeder layer did not improve larval growth, molting, or survival compared to that obtained with FeCS alone. The L4s derived from in vitro culture, when injected intraperitoneally into jirds, developed to sexually mature, microfilarial-producing adults in a normal in vivo time frame. Developing L4s isolated from jirds molted to young adults in vitro in NI medium plus serum.
采用体外培养方法研究马来布鲁线虫幼虫的特定营养需求。从埃及伊蚊（利物浦株）分离出的感染性三期幼虫（L3）在NCTC 135:IMDM（NI）培养基中添加不同类型血清，于5%二氧化碳/空气和5%二氧化碳/氮气环境下培养。幼虫生长、发育并蜕皮至四期（L4）；然而，L4中期之后的进一步生长和分化很有限。血清补充对于诱导马来布鲁线虫L3蜕皮是必要的。与所测试的其他4种血清相比，补铁小牛血清（FeCS）始终能促进更高比例的蜕皮。通过测试已知对体外培养的哺乳动物细胞或其他寄生虫生长重要的补充剂，评估了马来布鲁线虫幼虫的特定营养需求。与单独使用FeCS相比，添加不同浓度和组合的血红蛋白、胰岛素、转铁蛋白、硒、白蛋白、胆固醇或细胞饲养层，并未改善幼虫的生长、蜕皮或存活情况。体外培养获得的L4经腹腔注射到沙鼠体内后，在正常的体内时间框架内发育成性成熟、产微丝蚴的成虫。从沙鼠分离出的发育中的L4在添加血清的NI培养基中于体外蜕皮为年轻成虫。
