Acanthoeca spectabilis (ATCC® PRA-103)

Strain Designations: Ellis  /  Depositor: B Leadbeater  /  Biosafety Level: 1

Strain Designations Ellis
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

April 18, 1996 from salt marsh in Swan Bay, Queenscliffe, Australia.
Product Format frozen
Type Strain no
Phylogenetic analyses of SSU rRNA, LSU rRNA, tubA, and hsp-90
Medium ATCC® Medium 1525: Seawater 802 medium
Growth Conditions
Temperature: 20.0°C
Growth condition: monoxenic with Enterobacter aerogenes ATCC 13048
Protocol: Thaw the frozen material in a 35.0°C water bath and transfer the contents into a T25 flask containing 10 ml of growth medium. Flagellates should be observed within 4-6 days. Subculture on a weekly basis by scraping the bottom of the flask with a cell scraper and transferring one tenth of the culture into a new flask with medium.
Cryoprotective Solution

DMSO                                                                                  1.5 ml

Fresh growth medium w/o bacteria                               8.5 ml

1.     Mix the components in the order listed. When the medium is added to the DMSO the solution will warm up due to chemical heat.

2.     Harvest cells from a culture that is at or near peak density by filtration and centrifugation at 800 x g for 5 min.

3.     Adjust the concentration of cells at least 2 x 106/ml in fresh medium.

4.     Mix the cell preparation and the cryoprotective solution in equal portions.

5.     Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).

6.     Place vials in a controlled rate freezing unit. From room temperature cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  

7.     Ampules are stored in either the vapor or liquid phase of a nitrogen refrigerator.

8.     To establish a culture from the frozen state place the vial in a 35°C water bath.  Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.  Immediately after thawing, do not leave in water bath, aseptically remove the contents of the ampule and inoculate into a T-25 tissue culture flask containing 10 ml of an equal-parts mixture of ATCC media 1525 and 1405 bacterized with Klebsiella pneumoniae subsp. pneumoniae (ATCC® 700831) or Enterobacter aerogenes (ATCC® 13048)..

9.     Incubate at 20-25°C with the cap screwed on tightly.

10.   Once the culture is established, vigorously agitate the flask and aseptically transfer 0.5 ml to 10.0 ml of fresh bacterized medium.

11.   Follow the protocol for maintenance of culture.

Name of Depositor B Leadbeater

Leadbeater BS, et al. Choanoflagellate lorica construction and assembly: the nudiform condition. II. Acanthoeca spectabilis Ellis. Protist 159: 495-505, 2008. PubMed: 18485816

Leadbeater BS. Developmental and ultrastructural observations on two stalked marine choanoflagellates, Acanthoecopsis spiculifera Norris and Acanthoeca spectabilis Ellis. Proc. R. Soc. Lond. B Biol. Sci. 204: 57-66, 1979. PubMed: 37513

Carr M, et al. Molecular phylogeny of choanoflagellates, the sister group to Metazoa. Proc. Natl. Acad. Sci. USA. 105: 16641-16646, 2008. PubMed: 18922774