Home › Microbiology › Isolation and Characterization of Mesenchymal Stromal Cells from Human Umbilical Cord and Fetal Placenta
Microbiology JoVE (Open Access) Citable · DOI

Isolation and Characterization of Mesenchymal Stromal Cells from Human Umbilical Cord and Fetal Placenta

DOI: 10.3791/55224-v
What you'll learn
  • ✓ Dissect human umbilical cord and placental tissues into distinct anatomical regions
  • ✓ Isolate mesenchymal stromal cells using explant culture technique
  • ✓ Characterize MSC identity via colony-forming efficiency and marker expression analysis
Protocol

Here, we present a protocol for the dissection of human umbilical cord (UC) and fetal placenta sample into cord lining (CL), Wharton's jelly (WJ), cord-placenta junction (CPJ), and fetal placenta (FP) for the isolation and characterization of mesenchymal stromal cells (MSCs) using the explant culture technique.

Difficulty
intermediate
Total time
~7–10 days (tissue dissection ~2–3 hours; explant culture and characterization ~7–10 days)
Model organism
Human umbilical cord and fetal placenta (perinatal tissues)
Biosafety
BSL-1

Steps

1
Process and dissect umbilical cord and placental tissues

Dissect human umbilical cord (UC), cord-placenta junction (CPJ), and fetal placenta (FP) samples into anatomically distinct regions: cord lining (CL), Wharton's jelly (WJ), CPJ, and FP. Prepare tissue samples for cell isolation.

▶ 01:05
2
Isolate mesenchymal stromal cells via explant culture

Culture tissue explants using explant culture technique to allow MSCs to migrate and proliferate from the tissue fragments. Monitor culture conditions and cell outgrowth over days to weeks.

▶ 04:29
3
Characterize MSC markers and colony-forming efficiency

Assess isolated cells by measuring colony-forming efficiency and analyzing expression of mesenchymal stromal cell markers to confirm MSC identity and compare characteristics across perinatal tissue sources.

▶ 06:21
💬 Comments coming soon