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Cell Biology JoVE (Open Access) Citable · DOI

Culture of Murine Embryonic Metatarsals: A Physiological Model of Endochondral Ossification

DOI: 10.3791/54978-v
What you'll learn
  • ✓ Dissect embryonic murine metatarsals without contamination or tissue damage.
  • ✓ Culture ex vivo bone explants under physiological conditions.
  • ✓ Assess endochondral ossification using this explant model system.
  • ✓ Compare explant physiology to monolayer and 3D culture approaches.
Protocol

We present a protocol to dissect and culture embryonic day 15 (E15) murine metatarsal bones. This highly physiological ex vivo model of endochondral ossification provides conditions closer to the in vivo situation than cells in monolayer or 3D culture and is a vital tool for investigating bone growth and development.

Difficulty
advanced
Total time
~1–2 hours per litter (dissection + setup); culture maintained for 7–14 days
Model organism
Mouse (embryonic day 15)
Biosafety
BSL-1

Steps

1
Prepare dissection tools and workspace

Sterilize dissection instruments and arrange a clean workspace with appropriate lighting and magnification. Prepare culture media and reagents needed for metatarsal isolation.

▶ 00:51
2
Dissect and isolate embryonic metatarsals

Remove E15 embryos from timed-pregnant dams, carefully dissect forelimbs/hindlimbs under a dissecting microscope, and isolate individual metatarsal bones while preserving structural integrity.

▶ 01:47
3
Culture metatarsal explants in physiological conditions

Transfer isolated metatarsals to culture dishes with appropriate medium, maintaining oxygen and nutrient levels to recapitulate in vivo endochondral ossification.

▶ 04:57
4
Analyze bone growth and ossification outcomes

Monitor cultured metatarsals over time using imaging or histological analysis to measure endochondral ossification progression and validate model physiological relevance.

▶ 04:57
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