TGF-β1 negatively regulates cycling short- and long-term hematopoietic stem cells

Transforming growth factor-β (TGF-β) is considered to play a role in the maintenance of quiescent hematopoietic stem cells (HSCs) in vivo. We asked a question of whether TGF-β could be used to control the cell cycle status of HSCs in vitro. To examine the effect of TGF-β on the HSC function, we used in vitro culture system in which HSCs divide with retention of short- and long-term reconstitution ability. Single-cell analyses showed that regardless of its concentrations, TGF-β slowed down cell cycle progression of HSCs, but consequently suppressed the self-renewal potential, particularly in cycling HSCs. This study highlighted a role of TGF-β in the negative regulation of HSC number and function. Single-cell RNA-sequence analysis was performed on single cells isolated from the culture with SCF+TPO and SCF+TPO+TGF-β1 on day5. Seven or eight cells were isolated from individual wells with or without TGF-β1. A total of 135 single cells were randomly manipulated by mouth-pipette and transferred into cell lysis buffer. In the attached file "Sample_Barcode_Information.xlsx", the information of all sequenced cellls is listed in sheet1 and barcode information is listed in sheet2. Each cell is named as IVHSC_X(treatment)_X(batch information)_scX(barcode information, see sheet2)