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. 2011 Oct 21;44(1):26–35. doi: 10.3858/emm.2012.44.1.002

Figure 1.

Figure 1

Outline of the 3-stage differentiation protocol and stage-specific cell cluster morphology. Schematic representation of the 3-stage protocol to derive insulin-producing cells (IPCs) from hLMDFs. Stage 1: Expansion of hLMDFs. hLMDFs were isolated from labia minora dermis and placed in growth medium. Stage 2: MSC-endoderm transition stage. Cells were cultured in serum-free DMEM/F12 media supplemented with ITS for 1 week. During this stage, the cells metamorphosed into round epithelial-like cells. After 7 days of stage 2, the cells were found to express nestin by FACS analysis and to express endoderm markers by RT-PCR and western blot analysis. Stage 3: Differentiation of pancreatic islet-like cell clusters. Cells were cultured in stage 3 medium (DMEM + 10% FBS + ITS mix + nicotinamide). The cells continued to differentiate and form larger clusters. Essential factor manipulations at each stage are also shown. Scale bars, 200 µm.