UW-Madison researchers have developed a method for differentiation of human pluripotent stem cells (hPSCs) into endothelial cells (ECs) resulting in a model having improved fidelity to BBB gene expression and function. During the differentiation, cell populations are treated with small molecule CHIR99021 along with overexpression of one or a combination of key BBB transcription factors (TFs) DACH1, DACH2, FLI1, FOS, FOXC1, FOXF1, FOXF2, FOXQ1, HES1, JUN, KLF2, KLF4, LEF1, MECOM, NR4A1, NR4A2, PPARD, TBX3, TSC22D1, ZIC2 and ZIC3. Overexpression of one or a combination of these transcription factors induces global transcriptomic similarity to in vivo BBB ECs, including changes that reflect key BBB properties. These include increased tight junction protein expression (OCLN and CLDN5), increased transporter protein expression (MFSD2A, SLC2A1, SLC38A5, SLC7A5, ABCB1, and ABCG2) and reduced transcytosis-related protein expression (CAV1 and CAV2) and reduced leukocyte adhesion molecule expression (ICAM1).