Table 1.

Experimental Plan

Experimental conditions
	2D	3D
Experimental groups	Maintenance	Maintenance	Early differentiation	Late differentiation
Control	BM+LIF (3 days)	BM+LIF (3 days)	EDM+mSCF (8 days)	HDM (10 days)
CM1	BM+LIF (3 days)	BM+HepG2 CM (3 days)	EDM+mSCF (8 days)	HDM (10 days)
CM2	BM+LIF (3 days)	BM+HepG2 CM (3 days)		HDM (18 days)

Three mESCs groups were assessed: (1) control (3 days' standard maintenance medium and then 8 days' early differentiation medium), (2) CM1 (3 days' CM and then 8 days' early differentiation medium), and (3) CM2 (3 days' CM, bypassing early differentiation). All underwent hematopoietic differentiation with hEPO, mIL-3, and mSCF to 21 culture days.

2D, two-dimensional; 3D, three-dimensional; BM, basal medium; EDM, early differentiation medium; HDM, hematopoietic differentiation medium; LIF, leukemia inhibitory factor; mESC, murine embryonic stem cell; mSCF, murine stem cell factor.