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

Membranes

Ana Carolina Irioda¹  Bassam Felipe Mogharbel¹  Daiany de Souza Dobuchak¹  Katherine Athayde Teixeira de Carvalho¹  Priscila Elias Ferreira Stricker¹  Alyne Rodrigues de Araújo²  Celia Regina Cavichiolo Franco³  Felipe Azevedo Borges⁴  Rondinelli Donizetti Herculano⁴  Juan Carlos Chachques⁵  Carlos Frederico de Oliveira Graeff⁶  José Roberto de Souza Almeida Leite⁷ 

[1] Advanced Therapy and Cellular Biotechnology in Regenerative Medicine Department, Child and Adolescent Health Research and Pequeno Príncipe Faculties, Pelé Pequeno Príncipe Institute, Curitiba 80240-020, Brazil;Biodiversity and Biotechnology Research, Parnaíba Delta Federal University, Parnaíba 64202-020, Brazil;Cell Biology Department, Federal University of Paraná, Curitiba 81530-000, Brazil;Faculty of Pharmaceutics Sciences, São Paulo State University (UNESP), Araraquara 14800-903, Brazil;Laboratory Biosurgical Research, Cardiovascular Division, Pompidou Hospital, University of Paris, 75015 Paris, France;Physics Department, São Paulo State University (UNESP), Bauru 17033-360, Brazil;Research Center in Applied Morphology and Immunology, NuPMIA, Faculty of Medicine, University of Brasília, Brasília 70910-900, Brazil;

关键词: mesenchymal stem cells;    umbilical cord;    blood;    membrane;    biopolymer;    cholinergic neurons;   

DOI  :  10.3390/membranes11080598

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown