Differentiation of bone marrow cells into cells that express liver-specific genes in vitro: implication of the Notch signals in differentiation.
Differentiation of bone marrow cells into cells that express liver-specific genes in vitro: implication of the Notch signals in differentiation.
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DOI：
10.1016/s0006-291x(03)00637-5
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发表时间：
2003-05
影响因子：
3.1
通讯作者：
K. Okumoto;Takafumi Saito;Etsuko Hattori;Jun-itsu Ito;Tohru Adachi;T. Takeda;K. Sugahara;Hisayoshi Watanabe;Koji Saito;H. Togashi;S. Kawata
中科院分区：
文献类型：
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作者：
K. Okumoto;Takafumi Saito;Etsuko Hattori;Jun-itsu Ito;Tohru Adachi;T. Takeda;K. Sugahara;Hisayoshi Watanabe;Koji Saito;H. Togashi;S. Kawata
骨髓（BM）干细胞已被证明可分化为肝细胞。骨髓干细胞的分选和培养仍然很困难，这些细胞中肝脏特异性蛋白的基因表达尚未得到充分研究。我们使用负选择性磁性细胞分离系统来获得干细胞富集的BM细胞。将获得的细胞与肝细胞或肝细胞生长因子（HGF）一起培养，并通过逆转录-聚合酶链反应研究BM细胞向表达肝特异性基因、肝细胞核因子（HNF）1α、细胞角蛋白（CK）8、甲胎蛋白（AFP）和白蛋白的细胞的分化。我们还研究了Notch受体-1（Notch-1）及其配体Jagged-1在骨髓细胞分化中的基因表达。分选的骨髓细胞经免疫荧光染色显示Sca-1（Ataxin-1）阳性。荧光激活细胞分选仪分析显示，32.6%的分选的BM细胞具有造血干细胞标志物CD 90（Thy-1）的高水平表达。当与肝细胞一起培养时，这些细胞在培养第3天表达肝脏特异性基因HNF 1 α和CK 8，在培养第7天表达AFP和白蛋白。加入20 ng/ml HGF后，第3天细胞表达HNF 1 α，第7天细胞表达CK 8。在第3天，在培养的BM细胞中检测Notch-1和Jagged-1的基因表达。这些结果表明，负选择性磁性细胞分离系统是有用的干细胞富集的BM细胞的快速制备，并且Notch信号通路在体外BM细胞分化成肝细胞谱系中起作用。
Bone marrow (BM) stem cells have been shown to differentiate into liver cells. It remains difficult to sort and culture BM stem cells, and the gene expression of liver-specific proteins in these cells has not been fully investigated. We used a negative selective magnetic cell separation system to obtain stem cell-enriched BM cells. The cells obtained were cultured with hepatocytes or with hepatocyte growth factor (HGF), and the differentiation of BM cells into cells expressing liver-specific genes, hepatocyte nuclear factor (HNF) 1α, cytokeratin (CK) 8, α-fetoprotein (AFP), and albumin was investigated by the reverse transcription-polymerase chain reaction. We also investigated the gene expressions of Notch receptor-1 (Notch-1) and its ligand Jagged-1 in BM cell differentiation. Sorted BM cells showed positive for Sca-1 (Ataxin-1) by immunofluorescence staining. Fluorescence activated cell sorter analysis showed that 32.6% of sorted BM cells had a high level of expression of the hematopoietic stem cell marker CD90 (Thy-1). When cultured with hepatocytes, these cells expressed the liver-specific genes HNF1α and CK8 on culture day 3, AFP and albumin on culture day 7. When cultured with HGF (20ng/ml), the cells expressed HNF1α on day 3 and CK8 on day 7. Gene expressions of Notch-1 and Jagged-1 were detected in cultured BM cells on day 3. These results suggest that the negative selective magnetic cell separation system is useful for the rapid preparation of stem cell-enriched BM cells, and that the Notch signaling pathway plays a role in BM cell differentiation into a hepatocyte lineage in vitro.