Efficient One-Step Induction of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Produces MSC-Derived Neurospheres (MSC-NS) with Unique Transcriptional Profile and Enhanced Neurogenic and Angiogenic Secretomes

Figure 4

Transcriptional profiling analysis after neurobasal medium induction in UC-MSCs. (a) Results of reverse transcriptase PCR. Human NSCs (NouvNeu hNSC, NC0001) were used as a positive control. RT(-): no reverse transcriptase negative control. (b) Quantitative real-time PCR shows that neurosphere medium induction leads to significant overexpression of stemness and neurogenic genes as early as 24 hours after the induction. D0, UC-MSCs before induction; D1, 2, 3, 5, and 7 represent the RNA expression levels at Days 1, 2, 3, 5, and 7 after the induction. The expression of genes was normalized to that of GAPDH. Triplicate PCR amplifications were performed for each sample, and the results are represented as of three triplicate samples. (c) RNA sequencing analysis of UC-MSCs before and 48 hours after neurosphere formation. Heatmap represents supervised clustering analysis of transcriptional profiles which sharply distinguishes UC-MSCs (D001, D002, and D003) from UC-MSC-derived neurospheres (D21, D22, and D23) and human neural stem cells (NSC1 and NSC2). GO annotation map shows that genes involved with the regulation of nervous system development, regulation of neurogenesis, and stem cell population maintenance were significantly enriched after the induction.

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