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Long-term in vitro generation of amoebocytes from the Indian horseshoe crab Tachypleus gigas (Muller)

DRS at CSIR-National Institute of Oceanography

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Title Long-term in vitro generation of amoebocytes from the Indian horseshoe crab Tachypleus gigas (Muller)
 
Creator Joshi, B.
Chatterji, A.
Bhonde, R.
 
Description Amoebocyte is the single type of cell circulating in the horseshoe crab hemolymph, which plays a major role in the defense system of the animal. Granules present in these cells are sensitive to nanogram quantities of bacterial endotoxins, which form the basis of the Limulus amoebocyte lysate (LAL) test. Normally, amoebocytes for the production of the LAL are collected by cardiac puncture: hence, development of the in vitro culture system for amoebocytes will reduce the variability of the lysate and help to conserve the 400 million-yr-old living fossil. In the present investigation, organ culture of gill flaps have been attempted, that shown to be the source of amoebocytes. The gill flaps were cultured at 28 degrees C on a rocker platform in a modified L-15 medium supplemented with 10% v/v horseshoe crab serum. This led to the release of amoebocytes outside the gill flaps for a period of 6-8 wk with a more or less steady number of amoebocytes during the weekly harvest. No significant difference was seen in the yield of amoebocytes from male and female horseshoe crabs. Confocal laser microscopy studies revealed significant difference in the size of amoebocytes released in vitro as compared with those obtained in vivo. Thus, the culture conditions for the long-term generation of amoebocytes in vitro have been optimized from the Indian horseshoe crabs Tachypleus gigas by reducing the incidence of contamination, simulating in vivo conditions for the organ culture of gill flaps and improvising the nutritional status using the modified L-15 medium providing the desired osmolarity and pH
 
Date 2008-08-02T11:19:47Z
2008-08-02T11:19:47Z
2002
 
Type Journal Article
 
Identifier In Vitro Cellular & Developmental Biology - Animal. Vol. 38(5); 255-257p.
http://drs.nio.org/drs/handle/2264/1329
 
Language en
 
Publisher Springer