New method to differentiate human peripheral blood

monocytes into insulin producing cells

: Human hematosphere culture

Supplementary Figures

Supplementary Fig. 1. (Kim)

PBMC isolation & Generation of Hematosphere

Peripheral blood from healthy volunteer

Ficoll-Hypaque

Isolation of MNCs using Ficoll

3D cultureHigh cell density

Ultra low attach dish

Mononuclear cells(MNCs) layer

Culture of Blood derivedHematosphereEBM-2 (5%FBS)

Supplementary Fig 1. Generation of BBHS

(A) Schematic protocol for generation of hematosphere in human.

Unstained BBHS BBHS Stage-1

Stage-2 Stage-3 Stage-4

Supplementary Fig. 2. (Kim)

Supplementary Fig 2. Detection of insulin protein via DTZ staining

The intensity of insulin staining via DTZ (binding of zinc ion to insulin protein) increased gradually throughout the distinct four-stage of IPCs-development. Scale bar represents 10 ㎛