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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: In vivo conversion of rat astrocytes into neuronal cells through neural stem cells in injured spinal cord with a single zinc-finger transcription factor

Fig. 1

Reprogramming adult rat derived astrocytes. a A map of the lentiviral vectors, Sox2 and Zfp521. The vector is inducible by doxycycline (Dox) and contains the TRE element, a promoter, and Zfp521 or Sox2 CDS. b A summary of the experimental design for transdifferentiation of astrocytes into induced neural stem cells (iNSCs). Viral induction was performed at week 0. The cells were cultured in induction medium (IM) in the presence of Dox for 3 weeks with decreasing concentrations of fetal bovine serum (FBS) and increasing concentrations of growth factors (GF), bFGF, and EGF. The culture medium was then changed to neural stem cell medium (NSCM) supplemented with GF until week 4. The cells were transduced with Zfp521 (Zfp), Sox2, or both transcription factors (2TF). c qRT-PCR analysis of astrocyte (AST) markers (Gfap, S100β) and NSC markers (Sox2, Nestin, and Dcx) at 4 WPT. Data were normalized against GAPDH and presented relative to the expression of each indicated gene in astrocytes. d Immunofluorescent staining of cells for astrocytes and iNSC at 4 weeks post-transduction (WPT). Nuclei were counterstained with DAPI. e Quantification of protein markers revealed by immunostaining in d. Data in c and e are given as mean ± SD for three independent experiments. Data were analyzed by ANOVA and Tukey test as post hoc. *p < 0.05, **p < 0.01, and ***p < 0.001. The number of counted cells is presented in Additional file 4: Table S3

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