Collection and Transport at Cold Temperature of Cauda Epididymis

1. Male mouse (over 12 weeks old)
2. Fine scissors
3. Pair of watchmaker's #5 forceps
4. Wound clip (Autoclip 9mm; Clay Adams 427631) and clip applicator (Mik-Ron Autoclip Applier; Clay Adams 427630)
5. Temperature data logger (Thermochron iButton Cat.No.DS1921G; Maxim Integrated Products)
6. Preservation solution (KYUDO CO.,LTD. fertiupkyudo.co.jp)
7. CARD cold temperature transport kit (KYUDO CO.,LTD. fertiupkyudo.co.jp)
   • Thermos bottle (Cat.No.JMK-501: Thermos K.K.)
   • Paper box (in which a 0.2mL tube can stand)
   • Cotton wool
   • Cold packs (small and large)
   • Polystyrene foam transport box (KARUX KC-3)

1. Anesthetize a male mouse.
   
   
2. Make a small incision in the scrotum of the mouse and expel a cauda epididymis.
   
   
   
   
   
   
3. Cut the vas deferens and the corpus epididymis, and collect the cauda epididymis.
   
   
   
   
   
   
   [Collected Cauda Epididymis]
   
   
   
   
   
   
   
4. Push the testis back into the abdomen and close the wound using a wound clip.
   
   
5. Keep the mouse warm on a 37℃ warming plate until the mouse recovers from the effects of the anesthesia.
   

   Comment:

   1 week after the operation, the male mouse can be used for mating with a female mouse.

   
   

1. Put the removed cauda epididymis into the 0.2mL tube containing preservation solution.
   
   
   




2. Place the tube containing the cauda epididymis, a temperature data logger and a piece of cotton wool in the paper box.
   
   
   



3. Insert two cold packs (small) into a thermos bottle.
   
   
   
   
   
   
4. Insert the paper box containing the cauda epididymis into the thermos bottle.
   
   
   
   
   
   
5. Close the inner bottle cap.
   
   
   

   Note:

   When closing on the inner bottle cap, the paper box will fold inwards due to the bottle being packed with both cold packs and the paper box. Be sure not to place the paper box upside down; it is important to check the direction in which the paper box is lying through the slit of the inner bottle cap.

   
   
   
   
   
   
   
6. Close the outer bottle cap.
   
   
   
   
   
7. Place a cold pack (large) at the bottom of a foam transport box, then put the thermos bottle on top of it.
   
   
8. Pack one cold pack (large) on either side of the bottle, then set a further pack (large) on top and close the lid.
   

   Note:

   Take care not to place the paper box upside down.

   Note:

   It is only possible to place the thermos bottle in the center of the foam transport box and not the actual bottom, because the length of the thermos bottle is the same as that of the inner length of the foam transport box. This is to protect the thermos bottle during shipping.

   
   
9. Seal the lid of the foam transport box using packing tape.
   
   
   
   
   
   
10. Keep the foam transport box in the refrigerator until a courier comes to pick it up.
    
    
11. Send the samples via a regular courier service.

    Note:

    The sample must be transferred at a refrigerated temperature. Please ask the courier service directly about conditions during transport.

    Comment:

    Epididymal sperm at cold temperature maintain fertilizing ability for up to 72 hours.

Center for Animal Resources & Development,
Kumamoto University,
card3gpo.kumamoto-u.ac.jp

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