Basic research on treatment of liver failure and prevention against liver fibrosis by transplantation of MAPC-like stem cells established from bone marrow

骨髓MAPC样干细胞移植治疗肝衰竭和预防肝纤维化的基础研究

• 批准号: 18590269
• 负责人: MIYAZAKI Masahiro
• 金额: $ 2.47万
• 依托单位: Okayama Gakuin University (2007)Okayama University (2006)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590269/
• 关键词: a bone marrow-derived clonal mesenchymal stem cell; hepatocytes; transplantation; acute liver failure; liver fibrosis

Transplantation of hepatocytes or hepatocyte-like cells of extrahepatic origin is a promising strategy for treatment of acute and chronic liver failure. We examined possible utility of hepatocyte-like cells induced from bone marrow cells for such a purpose. We established a clonal cell line rBM25/S3 from rat bone marrow. The cells grew rapidly (doubling time,〜24 hours) without any appreciable changes in cell properties for at least 300 PDL over a period of 300 days, keeping normal diploid karyotype. The cells expressed CD29, CD44, CD49b, CD90, vimentin and fibronectin but not CD45, indicating that they are of mesenchymal cell origin. When plated on Matrigel with HGF and FGF-4, the cells efficiently differentiated into hepatocyte-like cells that expressed albumin, CYP1A1, CYP1A2, G6Pase, TO, TAT, HNFlα, and HNF4α. When cultivated in the same medium but on a collagen matrix, the cells prominently exhibited adipogenic properties, such as progressive accumulation of lipid droplets in the cytoplasm and expression of adiponectin, leptin and resistin.We intrasplenically transplanted the three types of cells into rats before 90% hepatectomy to induce fatal liver failure, or rats before and after initiation of CC1_4 treatment to induce liver fibrosis. Intrasplenic transplantation of hepatocyte-like cells rescued 5 out of 15 rats with,fatal liver failure. Furthermore, transplantation of undifferentiated rBM25/S3 cells most effectively suppressed liver fibrosis, followed by those of the adipogenic cells and the hepatogenic cells. Expression of MMP-2 and MMP-9 was also highest in undifferentiated rBM25/S3 cells.In conclusion, the bone marrow-derived clonal stem cell line (rBM25/S3) is suitable to treat liver fibrosis and the hepatogenically-differentiated derivative is potent to rescue liver failure. So we will further examine effects of combined transplantation of the different cell types of the same clonal origin on treatment of acute and chronic liver failure.

肝外来源的肝细胞或肝细胞样细胞的移植是治疗急性和慢性肝衰竭的有前途的策略。我们研究了从骨髓细胞诱导的肝细胞样细胞为此目的的可能用途。我们从大鼠骨髓中建立了克隆细胞系 rBM25/S3。细胞生长迅速（倍增时间约24小时），在300天内至少300 PDL，细胞特性没有任何明显变化，保持正常的二倍体核型。这些细胞表达CD29、CD44、CD49b、CD90、波形蛋白和纤连蛋白，但不表达CD45，表明它们是间充质细胞来源的。当铺在含有 HGF 和 FGF-4 的基质胶上时，细胞有效分化为表达白蛋白、CYP1A1、CYP1A2、G6Pase、TO、TAT、HNF1α 和 HNF4α 的肝细胞样细胞。当在相同的培养基中但在胶原基质上培养时，细胞显着表现出脂肪形成特性，例如细胞质中脂滴的逐渐积累以及脂联素、瘦素和抵抗素的表达。我们在90%肝切除术之前将这三种类型的细胞移植到大鼠体内以诱导致命性肝衰竭，或者在开始CC1_4治疗之前和之后的大鼠体内 从而诱发肝纤维化。肝细胞样细胞的脾内移植挽救了 15 只患有致命性肝功能衰竭的大鼠中的 5 只。此外，未分化rBM25/S3细胞的移植最有效地抑制肝纤维化，其次是脂肪形成细胞和肝形成细胞。 MMP-2和MMP-9在未分化的rBM25/S3细胞中的表达也最高。 总之，骨髓源性克隆干细胞系（rBM25/S3）适合治疗肝纤维化，肝源性分化的衍生物可有效挽救肝衰竭。因此，我们将进一步研究同一克隆来源的不同细胞类型联合移植治疗急慢性肝衰竭的效果。

项目成果

骨髄由来間葉系幹細胞移植によるラット肝線維化抑制機構の一部はMMP-9の作用に基づく

骨髓间充质干细胞移植抑制大鼠肝纤维化的机制部分基于MMP-9的作用

• 发表时间: 2007
• 作者: Miyazaki M;Hardjo M;Masaka T;Tomiyama K;Mahmut N;et. al.;ハルジョ マルハエン
• 通讯作者: ハルジョ マルハエン

Isolation of a bone marrow-derived stem cell line with high proliferation potential and its application for preventing acute fatal liver failure

• DOI: 10.1634/stemcells.2007-0078
• 发表时间: 2007-11-01
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Miyazaki, Masahiro;Hardjo, Marhaen;Huh, Nam-Ho
• 通讯作者: Huh, Nam-Ho

Rescue of 90% hepatectomized rats with acute liver failure by intrasplenic transplantation of hepatocytes derived from bone marrow stem cells.

救援％20of％2090％％20肝切除％20大鼠％20与％20急性％20肝脏％20失败％20by％20脾内％20移植％20of％20肝细胞％20衍生％20来自％20骨％20骨髓％20干％20细胞。

• 发表时间: 2006
• 作者: Miyazaki M;Hardjo M;Masaka T;Tomiyama K;Mahmut N;et. al.;ハルジョ マルハエン;Hardjo M;宮崎 正博;Miyazaki M;Masahiro Miyazaki;Miyazaki M
• 通讯作者: Miyazaki M

Rescue of 90%-hepatectomized rats with acute liver failure by intrasplenic transplantation of hepatocytes derived from a newly established mesenchymal stem cell line from rat bone marrow.

救援%20of%2090%-肝切除%20大鼠%20与%20急性%20肝脏%20失败%20by%20脾内%20移植%20of%20肝细胞%20衍生%20来自%20a%20新%20建立%20间充质%20干%20细胞%20系%20来自%20大鼠

• 发表时间: 2006
• 作者: ハルジョ;マルハエン;間阪 拓郎;Miyazaki M
• 通讯作者: Miyazaki M

Bone marrow-derived mesenchymal stem cells reduce liver fibrosis partly through action of MMP-9 in rats.

骨髓间充质干细胞部分通过 MMP-9 的作用减轻大鼠肝纤维化。

• 发表时间: 2007
• 作者: Miyazaki M;Hardjo M;Masaka T;Tomiyama K;Mahmut N;et. al.;ハルジョ マルハエン;Hardjo M
• 通讯作者: Hardjo M