[Study of in vitro expansion and differentiation into neuron-like cells of human umbilical cord blood mesenchymal stem cells].
OBJECTIVE
To explore the isolation, purification and expansion of human umbilical cord blood mesenchymal stem cells (MSCs) into neuron-like cells in vitro.


METHODS
Human 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.


RESULTS
The 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.


CONCLUSIONS
MSCs in human umbilical cord blood can expand in vitro and differentiate into non-mesenchymal cells.
Hou
Lingling
L
Chinese Academy of Military Medical Science, Beijing 100850, China.
Cao
Hua
H
Wei
Guorong
G
Bai
Cixian
C
Zhang
Yong
Y
Wu
Zuze
Z
Pei Xt
Xue tao
Xt
D015703:Antigens, CD / Q000032:analysis; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D002452:Cell Count; D002454:Cell Differentiation; D002455:Cell Division; D005312:Fetal Blood / Q000166:cytology*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000032:analysis; D008648:Mesoderm / Q000737:chemistry / Q000166:cytology*; D016900:Neurofilament Proteins / Q000032:analysis; D009474:Neurons / Q000166:cytology*; D010751:Phosphopyruvate Hydratase / Q000032:analysis; D011990:Receptors, Transferrin; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology*
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C068126:CD71 antigen; D019012:Integrin beta1; D016900:Neurofilament Proteins; D011990:Receptors, Transferrin; D010751:Phosphopyruvate Hydratase