Home › Cell Biology › Fate Mapping of Human Embryonic Stem Cells by Teratoma Formation
Cell Biology JoVE (Open Access) Citable · DOI

Fate Mapping of Human Embryonic Stem Cells by Teratoma Formation

DOI: 10.3791/2036-v
What you'll learn
  • ✓ Prepare hESC pellets for in vivo transplantation studies.
  • ✓ Perform renal capsule grafting to generate teratomas in mice.
  • ✓ Harvest, fix, and section teratoma tissue for histological analysis.
  • ✓ Interpret teratoma sections to assess hESC differentiation potential.
Protocol

Biopharma Insights Directed differentiation of hESCs into specific cells has generated much interest in regenerative medicine. We provide a concise, step-by-step protocol for determining the in vivo fate of selected hESCs that provides a valuable tool for characterizing tissue-specific reagents for cell-based therapy.

Difficulty
advanced
Total time
~4–6 weeks (including teratoma formation and tissue processing)
Model organism
Mouse (immunocompromised strain, unspecified)
Biosafety
BSL-1

Steps

1
Understand hESC fate mapping and teratoma formation

Review the biological rationale for using teratoma assays to assess pluripotency and in vivo differentiation capacity of hESCs in regenerative medicine applications.

▶ 00:56
2
Prepare hESC pellet for transplantation

Collect and concentrate hESCs into a compact pellet suitable for surgical grafting into the renal capsule.

▶ 01:44
3
Graft hESC pellet into mouse renal capsule

Perform surgical implantation of the hESC pellet under the renal capsule to allow in vivo teratoma formation over several weeks.

▶ 03:51
4
Harvest, fix, and section teratoma tissue

Extract the teratoma-bearing kidney, apply fixative, embed in paraffin, and cut histological sections for microscopic examination.

▶ 06:23
5
Interpret representative teratoma histology sections

Examine stained teratoma sections to identify differentiated tissues from all three germ layers, confirming pluripotency and characterizing differentiation patterns.

▶ 07:21
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