Directing human embryonic stem cells towards functional endothelial cells easily and without purification

Area of Science:

• Stem cell biology
• Developmental biology
• Cellular differentiation

Background:

• Hemangioblasts are limited sources for bi-potential cells.
• Previous methods for endothelial cell (EC) derivation from hemangioblasts had low efficiency due to cellular damage from enzymes and fluorescent activated cell sorting (FACS).

Purpose of the Study:

• To develop a method for deriving endothelial cells (ECs) from human embryonic stem cells (hESCs) that avoids damaging factors like enzymes and FACS.
• To achieve a high yield of functional ECs rapidly.

Main Methods:

• Human embryonic stem cells (hESCs) were dissociated using a mild protocol.
• Human embryoid bodies (hEBs) were formed under hemangioblast differentiation conditions.
• hEBs underwent a two-stage cytokine treatment, followed by culturing of adhesive cells.

Main Results:

• A high yield of endothelial cells (ECs) was obtained without enzyme usage or FACS.
• The derived ECs expressed key endothelial markers and demonstrated both in vitro and in vivo functionality.
• Confluent EC cultures were achieved within 4 days of hEB formation, significantly faster than prior methods.

Conclusions:

• The genetic changes for EC derivation occur before blast colony formation.
• Avoiding enzymes and FACS prevents cell damage, leading to efficient EC derivation.
• This protocol offers a faster and gentler method for generating functional ECs from hESCs.