expression of pluripotency genes and neuronal markers after neurodifferentiation in fibroblasts co-cultured with human umbilical cord blood mononuclear cells

• Published in: In vitro cellular & developmental biology. Animal 2015-01, Vol.51 (1), p.26-35
• Main Authors: Marinowic, D. R, Domingues, M. F, Machado, D. C, DaCosta, J. C
• Format: Article
• Language: English
• Subjects: absorbance; adults; Animal Genetics and Genomics; Animals; Biomarkers - metabolism; Biomedical and Life Sciences; BIOTECHNOLOGY; blood; bone formation; Cell Biology; Cell Culture; Cell Differentiation - genetics; Cell Proliferation; coculture; Coculture Techniques - methods; cultured cells; Developmental Biology; fibroblasts; Fibroblasts - cytology; Fluorescent Antibody Technique; Gene Expression Regulation; genes; genetic markers; Humans; Imaging, Three-Dimensional; Interleukin-2 - metabolism; Leukocytes, Mononuclear - cytology; Leukocytes, Mononuclear - metabolism; Life Sciences; Mesoderm - cytology; Mice; microscopy; Neurons - cytology; Neurons - metabolism; NIH 3T3 Cells; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; reverse transcriptase polymerase chain reaction; Stem Cells; umbilical cord; Umbilical Cord - cytology
• ISSN: 1071-2690; 1543-706X
• DOI: 10.1007/s11626-014-9804-8

Human umbilical cord blood is an attractive source of stem cells; however, it has a heterogeneous cell population with few mesenchymal stem cells. Cell reprogramming induced by different methodologies can confer pluripotency to differentiated adult cells. The objective of this study was to evaluate the reprogramming of fibroblasts and their subsequent neural differentiation after co-culture with umbilical cord blood mononuclear cells. Cells were obtained from four human umbilical cords. The mononuclear cells were cultured for 7 d and subsequently co-cultured with mouse fibroblast NIH-3T3 cells for 6 d. The pluripotency of the cells was evaluated by RT-PCR using primers specific for pluripotency marker genes. The pluripotency was also confirmed by adipogenic and osteogenic differentiation. Neural differentiation of the reprogrammed cells was evaluated by immunofluorescence. All co-cultured cells showed adipogenic and osteogenic differentiation capacity. After co-cultivation, cells expressed the pluripotency gene KLF4. Statistically significant differences in cell area, diameter, optical density, and fractal dimension were observed by confocal microscopy in the neurally differentiated cells. Contact in the form of co-cultivation of fibroblasts with umbilical cord blood mononuclear fraction for 6 d promoted the reprogramming of these cells, allowing the later induction of neural differentiation.