Additional Supporting Information may be found in the online version of this article.

STEM_1052_sm_SuppFig1.pdf284KSupplementary Figure 1. Expression profiles of cell-surface markers in hUCBMSCs and hUCB-MSC-derived spheres (A) Flow cytometry analysis was performed using monoclonal antibodies directed against the mesenchymal stem cell-positive surface markers CD44 (upper panel) and CD90 (lower panel). (B) The expression patterns of CD34 and CD117, which are hematopoietic cell-specific markers, were assessed in monolayer-cultured MSCs and MSC-derived spheres using flow cytometry.
STEM_1052_sm_SuppFig2.tif1661KSupplementary Figure 2. Expression levels of adipogenic and osteogenic markers in MSC monolayers and spheres (A) The efficiency of adipogenic differentiation was evaluated through the expression of adipogenic marker genes, including C/EBP-β, aP2, PPARα and leptin. (B) Expression levels of the osteoblast-specific transcription factor RUNX2 and bonespecific osteocalcin were assessed to determine the efficiency of osteogenic induction.
STEM_1052_sm_SuppFig3.pdf220KSupplementary Figure 3. Expression profiles of integrin-related cell-surface markers in MSC monolayers and spheres The expression levels of CD49a, CD49b, CD49e, and CD104 were assessed using flow cytometry analysis of sphere-derived hMSCs and monolayer-cultured hMSCs. The results were charted after three individual flow cytometry analyses were performed. (*, P < 0.05; **, P < 0.01)
STEM_1052_sm_SuppFig4.pdf265KSupplementary Figure 4. Significance of p53 on the formation of spheres and the differentiation potential of hMSCs The hMSCs were analyzed after transfection with either control siRNA (siCTL) or the siRNA targeting p53 (sip53). (A) The expression levels of p53 were analyzed using RT-PCR 3 days after siRNA transfection. (B) After siRNA transfection, hMSCs were stained with propidium iodide, and their cell cycle distribution was determined using flow cytometry (n = 3). (C-D) After siRNA transfection, hMSCs were subjected to sphere-forming conditions. The sphere-forming efficiency (C) and sphere size (D) of each population were assessed. (**, P < 0.01, n = 50) (E) The efficiency of osteogenic (E) and adipogenic (F) differentiation was evaluated through the expression of osteogenic and adipogenic marker genes, respectively. β-actin was used for the loading control.
STEM_1052_sm_SuppFig5.pdf332KSupplementary Figure 5. Schematic diagram of the role of CD49f in MSC sphere formation and reprogramming This schematic summarizes the proposed role of CD49f in hUCB-MSCs. This diagram illustrates how the activation of CD49f can lead to the phosphorylation of PI3K/AKT, which in turn leads to an enhancement of MSC sphere formation and hMSC cellular potency. Additionally, CD49f expression can contribute to the maintenance of stem cell pluripotency.
STEM_1052_sm_SuppInfo.pdf136KSupplementary Data
STEM_1052_sm_SuppTab1.pdf112KSupplementary Table 1.

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