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Umbilical Mononuclear Cells and Fibroblast Interaction Downregulate the Expression of Cell Cycle Negative Control Genes

Author:
Marinowic, Daniel Rodrigo, Zanirati, Gabriele, Azevedo, Pamella Nunes, De Souza, Eduardo Vieira, Bruzzo, Fernanda, Silva, Samara Pedroso da, Heuser, Eliete Biasotto, Machado, Denise Cantarelli, Da Costa, Jaderson Costa
Source:
Cellular reprogramming 2018 v.20 no.5 pp. 320-327
ISSN:
2152-4998
Subject:
adult stem cells, blood, bone marrow, cell cycle, cell viability, coculture, fibroblasts, gene expression, gene expression regulation, genes, humans, messenger RNA, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, therapeutics, toxicity testing, umbilical cord
Abstract:
The human umbilical cord blood (HUCB) is an excellent source of adult stem cells, having the benefit of being younger than the bone marrow stem cells. The role of stem cells in the lesion repair mechanism is still being studied. We evaluated the capability of HUCB to interfere into the fibroblast dedifferentiation plasticity through cocultivation. Direct and indirect cocultures were maintained for 24, 48, and 72 hours. Coculture viability was evaluated by MTT assay. The messenger RNA was extracted, and the expression of p16 and p21 genes was estimated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). The direct or indirect contact did not interfere with fibroblast cell viability. However, these direct and indirect contacts reduced the expression of p16 and p21 genes. A sigmoidal curve was applied to adjust gene expression against time, and a mathematical function was established for gene expression according to cell culture type. These results suggest that the differentiated cells were influenced by immature cells (HUCB) either by the direct contact or by signaling molecules, which alter their behavior and plasticity. Therefore our data may contribute to paracrine effects other than the commonly known to be responsible for the repair of lesions in stem cell therapy.
Agid:
6446907