Ma Lian, Cui Bing-lin, Feng Xue-yong, Law Frie-da, Jiang Xue-wu, Yang Li-ye, Xie Qing-dong, Huang Tian-hua
Department of Pediatrics, 2nd Affiliated Hospital of Shantou University Medical College, Shantou 515041, China.
Zhonghua Er Ke Za Zhi. 2006 Jul;44(7):513-7.
To investigate the isolation and expansion of mesenchymal stem cells (MSCs) from human umbilical cord Wharton's jelly and their biological identities, and explore the possibility of inducing human umbilical cord-derived MSCs to differentiate into neurocyte-like cells.
The growth and proliferative abilities of human umbilical cord-derived MSCs were observed, and their immunophenotypes were determined by flow cytometry. Salvia miltiorrhiza and beta-sulfhydryl alcohol were adopted to induce the cells to differentiate. The differentiated and undifferentiated cells were identified with immunocytochemistry. The pleiotrophin and nestin genes were measured by RT-PCR.
A population of human umbilical cord-derived MSCs were isolated from human umbilical Wharton's jelly; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. The human umbilical cord-derived MSCs were positive for CD(29), CD(44), CD(59), CD(105), but negative or weakly expressed the markers of hematopoietic cells such as CD(14), CD(33), CD(34), CD(28), CD(45) and CD(117). The important GVHD correlation markers were negative or weakly expressed, including CD(80) (B7-1), CD(86) (B7-2), CD(40) and CD(40L). Salvia miltiorrhiza beta-sulfhydryl alcohol could induce the MSCs to express nestin, a marker of neuronal precursor stem cells at early stage of differentiation. Later, they exhibited neural phenotypes, expressing beta-tubulin III and neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that the MSCs could express pleiotrophin either before or after the induction of salvia miltiorrhiza, furthermore, after the induction the expression was markedly enhanced and the nestin gene was also expressed.
The human MSCs could be isolated from human umbilical cord Wharton's jelly, and it was easy to propagate these MSCs. The negative GVHD correlated markers might result from the fact that MSCs had no HLA barrier, which may suggest potential clinical significance. The MSCs are capable of differentiating into neurocyte-like cells and they may represent an alternative stem cell source for CNS cells transplantation.
研究从人脐带华通氏胶中分离和扩增间充质干细胞（MSCs）及其生物学特性，并探讨诱导人脐带源MSCs分化为神经样细胞的可能性。
观察人脐带源MSCs的生长和增殖能力，采用流式细胞术检测其免疫表型。采用丹参和β-巯基乙醇诱导细胞分化。用免疫细胞化学法鉴定分化和未分化细胞。用RT-PCR检测多效生长因子和巢蛋白基因。
从人脐带华通氏胶中分离出一群人脐带源MSCs；将其处理后获得成纤维细胞样细胞群体，可在体外长期维持，群体倍增稳定，在培养中作为未分化细胞传代超过10代，表明其增殖能力。人脐带源MSCs对CD(29)、CD(44)、CD(59)、CD(105)呈阳性，但对造血细胞标志物如CD(14)、CD(33)、CD(34)、CD(28)、CD(45)和CD(117)呈阴性或弱表达。重要的移植物抗宿主病（GVHD）相关标志物呈阴性或弱表达，包括CD(80)（B7-1）、CD(86)（B7-2）、CD(40)和CD(40L)。丹参β-巯基乙醇可诱导MSCs表达巢蛋白，巢蛋白是神经元前体细胞分化早期的标志物。随后，它们表现出神经表型，表达β-微管蛋白III、神经丝（NF）和胶质纤维酸性蛋白（GFAP）。RT-PCR证实，丹参诱导前后MSCs均可表达多效生长因子，诱导后表达明显增强，巢蛋白基因也表达。
人MSCs可从人脐带华通氏胶中分离得到，且易于扩增。GVHD相关标志物阴性可能是因为MSCs无HLA屏障，这可能具有潜在的临床意义。MSCs能够分化为神经样细胞，可能是中枢神经系统细胞移植的另一种干细胞来源。
