Additional file 1 of Ficoll density gradient sedimentation isolation of pelage hair follicle mesenchymal stem cells from adult mouse back skin: a novel method for hair follicle mesenchymal stem cells isolation
Creators: name: Gan, Yuyang
name: Wang, Hailin
name: Du, Lijuan
name: Fan, Zhexiang
name: Sun, Pingping
name: Li, Kaitao
name: Qu, Qian
name: Wang, Jin
name: Chen, Ruosi
name: Hu, Zhiqi
name: Miao, Yong
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: Cell Biology
subject: Genetics
subject: Molecular Biology
subject: Physiology
subject: Environmental Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Mathematical Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer
subject: Hematology

description: Additional file 1: Fig. S1. Schematic diagram of Pelage DPCs isolation. Fig. S2. Morphology, adhesion and cell migration of dermal papilla cells. Whisker (A) and Pelage (B) follicles morphology observed under stereoscope, no blood sinus was found in pelage follicle, the number of pelage follicles was much more than whisker follicles. Hair shafts and hair bulb located in the same suspensions after enzyme digestion and scarping (C). After two-steps isolation, several Pelage DP spheres can be seen under microscope (D-F, Red arrow). Whisker DP spheres were observed under microscope (G, white arrow). Pelage DPCs started emigration in 24 h (H, white arrow showed DPCs started emigrated out of the DP sphere) and proliferation, most of pelage DPCs finished emigration in 4 days (I, white arrow showed plenty of DPCs emigrated out of the DP sphere), while whisker DPCs started emigrate in 72 h and finished in 7 days (J). Pelage DPCs (K) and whisker DPCs (L) showed similar cell morphology and aggregation growth pattern. Fig. S3. Differential expression of Sox2 between whisker and pelage DPCs. Sox2 expression was detected in primary DP sphere and cultured DPCs. Whisker DP showed high Sox2 expression both in DP sphere and DPCs, while Pelage DP showed almost no expression on Sox2 whether in primary DP sphere or cultured DPCs. Fig. S4. Isolation and identification of Pelage HF-MSC and whisker HF-MSC exosomes. The morphology of exosomes was observed by electron microscope; Western Blotting verification specific markers CD9 and CD81; The detection particle size was between 30 and 100 nm. Fig. S5. Effect and distribution of Whisker HF-MSC treatment on dorsal hair growth. 3-week-old C57BL/6 mice were divided into Pelage HF-MSC group and control group. Pelage HF-MSC group enter into anagen period earlier than control group, which was similar to Pelage HF-MSC group. Labeling whisker HF-MSC with DIO (green), showed that whisker HF-MSC located around the host hair follicle (White arrow). Fig. S6. Effect and distribution of whisker HF-MSC Exosomes treatment on dorsal hair growth. 7-week-old C57BL/6 mice were divided into whisker HF-MSC exosomes group and control group (The same as the control group in Fig. 6). Exosomes were injected every other day, a total of three times injection were performed. On day 8, whisker HF-MSC exosomes group enter into anagen period earlier than control group. Injected whisker HF-MSC exosomes were labeled with DIO (green) and located around host hair follicle (white arrow). Fig. S7. Pictorial diagram to show the location of MSCs and exosomes injection protocol. The cell/exosomes suspension was injected along the midline of the back of the mouse, and the amount of cell suspension was 300 µl. HF-MSCs suspension injection was performed once, exosomes suspension injection was performed.