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Cryopreservation of infective-stage larvae of Brugia malayi
1Guizhou Provincal Institute of Parasitic Diseases, Guiyang, People's Republic of China.
Abstract:
Third-stage larvae of Brugia malayi (Guizhou, China strain), in RPMI-1640, 6% DMSO and 15% calf serum, frozen slowly in the vapor phase of liquid nitrogen prior to emersion in liquid nitrogen, remained viable for as long as as 321 days and were infective to Mongolian jirds. All DMSO should be removed from the freezing medium to restore larval activity after freezing.
Insights
Cryopreserved Brugia malayi larvae remain viable and infective for over 300 days. Removing dimethyl sulfoxide (DMSO) from the freezing medium is crucial for restoring larval activity post-thaw.
Area of Science:
- Parasitology
- Cryobiology
Background:
- Brugia malayi is a filarial nematode responsible for lymphatic filariasis.
- Effective cryopreservation methods are essential for maintaining parasite viability for research and control programs.
Purpose of the Study:
- To evaluate the long-term viability and infectivity of cryopreserved Brugia malayi larvae.
- To determine the optimal conditions for cryopreservation and post-thaw recovery.
Main Methods:
- Third-stage larvae of Brugia malayi were cryopreserved using a slow-freezing method in RPMI-1640 with 6% DMSO and 15% calf serum.
- Larval viability and infectivity were assessed after 321 days of cryopreservation.
- The effect of DMSO removal on larval activity was investigated.
Main Results:
- Cryopreserved Brugia malayi larvae remained viable and infective to Mongolian jirds for up to 321 days.
- Complete removal of DMSO from the freezing medium was necessary to restore larval activity after thawing.
Conclusions:
- Long-term cryopreservation of Brugia malayi larvae is feasible.
- DMSO removal is a critical step for successful post-thaw recovery of larval infectivity.
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