Cell Biology JoVE (Open Access) Citable · DOI

Culture and Maintenance of Human Embryonic Stem Cells

Name: Culture and Maintenance of Human Embryonic Stem Cells
Uploaded: 2026-06-17
Duration: 9 min 36 s
Description: Human embryonic stem (hES) cells must be monitored and cared for in order to maintain healthy, undifferentiated cultures. At minimum, the cultures must be ...

DOI: 10.3791/1427-v

What you'll learn

✓ Maintain undifferentiated human embryonic stem cells in culture
✓ Apply proper passaging and feeding protocols for hES cells
✓ Identify morphological markers of healthy, pluripotent hES cell colonies
✓ Evaluate cell viability and contamination risk during routine maintenance

Protocol

This protocol demonstrates how to maintain healthy, undifferentiated human embryonic stem (ES) cells.

Difficulty

intermediate

Total time

~30 min per passage (routine maintenance); ~2–3 days between passages

Model organism

Human embryonic stem cells (hES)

Biosafety

BSL-1

Steps

Understand hES cell culture fundamentals

Review the biological requirements and culture conditions necessary to maintain human embryonic stem cells in an undifferentiated, pluripotent state.

▶ 00:07

Perform routine hES cell maintenance protocols

Execute standard culture techniques including medium changes, cell observation, passaging, and colony management to sustain healthy undifferentiated hES cell populations over multiple culture cycles.

▶ 00:25

Evaluate and document representative results

Assess morphology, viability, and pluripotency markers of maintained hES cells; document observations to confirm protocol success and cell line integrity.

▶ 08:24

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