OBJECTIVETo explore the isolation, purification and expansion of human umbilical cord blood mesenchymal stem cells (MSCs) into neuron-like cells in vitro.

METHODSHuman cord blood samples were obtained sterilely with 20 U/ml preservative-free heparin. MSCs were isolated by lymphocyte separation medium (density 1.077 g/ml), and purified and expanded with Mesencult trade mark medium. The surface antigen expression of MSCs was detected by flow cytometry. The passage 2, 5 and 8 of the expanded MSCs were induced to differentiate to neuron-like cells. Specific markers and structures were detected by immunohistochemistry and histochemistry methods.

RESULTSThe number of MSCs increased two- to three-fold with each expanded passage. 6.6 x 10(5) primary MSCs were expanded ten passages to reach a number of 9.9 x 10(8), and was increased about 1.5 x 10(3)-fold. Flow cytometry showed that MSCs did not express antigens CD(34), CD(11a) and CD(11b), but expressed strongly CD(29) and weakly CD(71), which was identical to human bone marrow-derived MSCs. 70% cells exhibited typical neuron-like phenotype after induction. Immunohistochemistry staining showed that all of the induced different-passage MSCs expressed neurofilament (NF) and neuron-specific enolase (NSE). Special Nissl body was found by histochemistry.

CONCLUSIONMSCs in human umbilical cord blood can expand in vitro and differentiate into non-mesenchymal cells.

Antigens, CD ; analysis ; Antigens, Differentiation, B-Lymphocyte ; analysis ; Cell Count ; Cell Differentiation ; Cell Division ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Mesoderm ; chemistry ; cytology ; Neurofilament Proteins ; analysis ; Neurons ; cytology ; Phosphopyruvate Hydratase ; analysis ; Receptors, Transferrin ; Stem Cells ; chemistry ; cytology