Hou Lingling, Cao Hua, Wang Dongmei, Wei Guorong, Bai Cixian, Zhang Yong, Pei Xuetao
Beijing Institute of Transfusion Medicine, Beijing, China.
Int J Hematol. 2003 Oct;78(3):256-61. doi: 10.1007/BF02983804.
Mesenchymal stem cells (MSCs) in human umbilical cord blood are multipotent stem cells that differ from hematopoietic stem cells. They can differentiate in vitro into mesenchymal cells such as osteoblasts and adipocytes. However, differentiation into nonmesenchymal cells has not been demonstrated. Here, we report the isolation, purification, expansion, and differentiation of human umbilical cord blood MSCs into neurocytes in vitro. Cord blood samples were allowed to drain from the end of the cord into glass bottles with 20 U/mL preservative-free heparin. MSCs were isolated from human umbilical cord blood, purified, and expanded in Mesencult medium. Surface antigens of MSCs were analyzed by fluorescence-activated cell sorting (FACS). MSC passages 2,5, and 8 were induced to differentiate into neuron-like cells. Neurofilament (NF) and neuron-specific enolase (NSE) were detected by immunohistochemistry staining. Special Nissl bodies were observed by histochemical analysis. The results showed that 6.6 x 10(5) primary MSCs were expanded for 10 passages to obtain 9.9 x 10(8) MSCs, an increase of approximately 1.5 x 10(3)-fold. FACS results showed that the MSCs did not express antigens CD34, CD11a, and CD11b and expressed CD29 and CD71, an expression pattern identical to that of human bone marrow-derived MSCs. Induction results indicated that approximately 70% of the cells exhibited a typical neuron-like phenotype. Immunohistochemistry staining suggested that induced MSCs of different passages expressed NF and NSE. Special Nissl bodies were obvious in the neuron-like cells. These results suggest that MSCs in human umbilical cord blood are capable of differentiating into neuron-like cells in vitro.
人脐带血中的间充质干细胞（MSCs）是不同于造血干细胞的多能干细胞。它们在体外可分化为成骨细胞和脂肪细胞等间充质细胞。然而，向非间充质细胞的分化尚未得到证实。在此，我们报告了人脐带血MSCs在体外的分离、纯化、扩增以及向神经细胞的分化。将脐带血样本从脐带末端引流到含有20 U/mL无防腐剂肝素的玻璃瓶中。从人脐带血中分离出MSCs，在Mesencult培养基中进行纯化和扩增。通过荧光激活细胞分选（FACS）分析MSCs的表面抗原。诱导第2、5和8代的MSCs分化为神经元样细胞。通过免疫组织化学染色检测神经丝（NF）和神经元特异性烯醇化酶（NSE）。通过组织化学分析观察特殊的尼氏体。结果显示，6.6×10⁵个原代MSCs扩增10代后获得9.9×10⁸个MSCs，增加了约1.5×10³倍。FACS结果显示，这些MSCs不表达抗原CD34、CD11a和CD11b，表达CD29和CD71，其表达模式与人类骨髓来源的MSCs相同。诱导结果表明，约70%的细胞呈现典型的神经元样表型。免疫组织化学染色表明，不同代次的诱导MSCs均表达NF和NSE。在神经元样细胞中特殊的尼氏体明显。这些结果表明，人脐带血中的MSCs在体外能够分化为神经元样细胞。
