Human Mesenchymal Stem Cells Seeded on the Natural Membrane to Neurospheres for Cholinergic-like Neurons

• Published in: Membranes (Basel) Vol. 11; no. 8; p. 598
• Main Authors: Stricker, Priscila, de Souza Dobuchak, Daiany, Irioda, Ana, Mogharbel, Bassam, Franco, Celia, de Souza Almeida Leite, José, de Araújo, Alyne, Borges, Felipe, Herculano, Rondinelli, de Oliveira Graeff, Carlos, Chachques, Juan, de Carvalho, Katherine
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 07-08-2021; MDPI
• Subjects: Acetyltransferase; Alzheimer's disease; Antibiotics; biopolymer; Biopolymers; blood; CD105 antigen; CD34 antigen; CD45 antigen; CD73 antigen; CD90 antigen; Cell culture; Cell differentiation; Choline; Choline O-acetyltransferase; cholinergic neurons; Cholinergics; Differentiation; Ethics; Flow cytometry; Glial fibrillary acidic protein; Growth factors; Immunocytochemistry; Leukocytes (mononuclear); membrane; Membranes; Mesenchymal stem cells; Methods; Nestin; Neurons; Neurospheres; Phenotypes; Polymerase chain reaction; Precursors; Responsible persons; Stem cells; Transfection; Tubulin; Umbilical cord; St Louis Missouri; United States > US
• ISSN: 2077-0375; 2077-0375
• DOI: 10.3390/membranes11080598

This study aimed to differentiate human mesenchymal stem cells (hMSCs) from the human umbilical cord in cholinergic-like neurons using a natural membrane. The isolation of hMSCs from Wharton’s jelly (WJ) was carried out using “explant” and mononuclear cells by the density gradient from umbilical blood and characterized by flow cytometry. hMSCs were seeded in a natural functional biopolymer membrane to produce neurospheres. RT-PCR was performed on hMSCs and neurospheres derived from the umbilical cord. Neural precursor cells were subjected to a standard cholinergic-like neuron differentiation protocol. Dissociated neurospheres, neural precursor cells, and cholinergic-like neurons were characterized by immunocytochemistry. hMSCs were CD73+, CD90+, CD105+, CD34- and CD45- and demonstrated the trilineage differentiation. Neurospheres and their isolated cells were nestin-positive and expressed NESTIN, MAP2, ßIII-TUBULIN, GFAP genes. Neural precursor cells that were differentiated in cholinergic-like neurons expressed ßIII-TUBULIN protein and choline acetyltransferase enzyme. hMSCs seeded on the natural membrane can differentiate into neurospheres, obtaining neural precursor cells without growth factors or gene transfection before cholinergic phenotype differentiation.