| Affiliation: | (1) Department of Fisheries and Wildlife Sciences, Virginia Polytechnic Institute and State University, 24061 Blacksburg, Virginia;(2) Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 24061 Blacksburg, Virginia;(3) Horseshoe Crab Research Center, Virginia Polytechnic Institute and State University, 24061 Blacksburg, Virginia;(4) National Marine Fisheries Service RTR Unit at Virginia Tech, 100 Cheatham Hall, 24061-0321 Blacksburg, Virginia |
| Abstract: | Summary This study provides information relevant to future research aimed at producing Limulus Amebocyte Lysate (LAL) in vitro, which would potentially reduce the need to harvest and bleed horseshoe crabs as in the current methods of LAL production. To address the need for primary culture of horseshoe crab amebocytes, this study tested the effects of a variety of standard insect cell culture media on amebocyte morphology and viability after 7 d of maintenance. Amerbocyte morphology was least altered from in vivo form in Grace’s Modified Insect Medium, with no observed degranulation of cells, as compared to the other media tested. There were significant differences in amebocyte viability among the six insect cell culture media tested. Grace’s Modified Insect Medium sustained viability of 77.2±5.1% (mean ± standard deviation) of amebocytes, followed distantly by Grace’s Insect Medium with 35.1±8.7% amebocyte viability. Results indicate that Grace’s Modified Insect Medium with horseshoe crab serum supplementation was the best candidate of the six media tested for future medium optimization for Limulus amebocyte requirements. |
