Culture of aerobic bacteria from mouse mLN

Laboratorio de Inmunometabolismo

CULTIVO CELULAR
Autor/a

omclab

Fecha de publicación

13 de junio de 2026

Resumen

Extraer células linfoides de pulmón de ratón para los fines que se requiera.

  1. Euthanize the mouse.
  2. Open the peritoneal cavity and expose the intestines. See note.
  3. Carefully excise the mLN, removing the surrounding fat.
  4. Place the mLN in a sterile 40µm cell strainer (BD 352340), and place the strainer in a sterile 100mm petri dish (BD 351029).
  5. Add 500µl of sterile PBS (Ca2+/Mg2+ free) to the mLN.
  6. Using the flat top portion of a 3ml syringe plunger (BD 309585), mash the tissue to obtain a homogenate suspension.
  7. Collect the suspension from the petri dish and prepare serial dilutions (usually 1x10-1 to 1x10-3).
  8. Plate 200µl of each dilution in a blood agar plate (Tryptic soy agar with 5% sheep blood) for 24h at 37ºC.
  9. Proceed to DNA isolation. Note: Steps 2 to 8 are to be performed next to a Bunsen burner, or under a hood.

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