Optimal conditions for in vivo induction of dopaminergic neurons from embryonic stem cells through stromal cell-derived inducing activity
Optimal conditions for in vivo induction of dopaminergic neurons from embryonic stem cells through stromal cell-derived inducing activity
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DOI：
10.1002/jnr.10363
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发表时间：
2002-09-15
影响因子：
4.2
通讯作者：
Hashimoto, N
中科院分区：
文献类型：
--
作者：
Morizane, A;Takahashi, J;Hashimoto, N
利用基质细胞源性诱导活性（SDIA）诱导小鼠胚胎干细胞（ES）多巴胺（DA）神经元的方法已有报道。移植后，sdia诱导的DA神经元融入小鼠纹状体，并保持酪氨酸羟化酶（TH）表达阳性。在本研究中，为了优化移植效率，我们用SDIA处理小鼠ES细胞不同天数（8-14天）。用木瓜蛋白酶分离sdia处理的ES细胞菌落，然后移植到6-羟多巴胺（6-OHDA）损伤的小鼠纹状体中。胚胎干细胞移植前经SDIA处理12 d， th阳性存活细胞数与移植细胞总数之比最高。这一比例表明，移植细胞菌落比移植细胞悬液更有效地获得th阳性细胞。当我们将2 × 10(5)、2 × 10(4)或2 × 10(3)细胞的细胞悬液移植到6- ohda损伤的小鼠纹状体中时，我们观察到只有少数th阳性细胞存活。综上所述，通过SDIA诱导小鼠ES细胞DA神经元12 d，并将细胞集落移植到小鼠纹状体中是th阳性神经元在体内存活的最有效方法。(C) 2002 Wiley-Liss, Inc。
A method of inducing dopamine (DA) neurons from mouse embryonic stem (ES) cells by stromal cell-derived inducing activity (SDIA) was previously reported. When transplanted, SDIA-induced DA neurons integrate into the mouse striatum and remain positive for tyrosine hydroxylase (TH) expression. In the present study, to optimize the transplantation efficiency, we treated mouse ES cells with SDIA for various numbers of days (8-14 days). SDIA-treated ES cell colonies were isolated by papain treatment and then grafted into the 6-hydroxydopamine (6-OHDA)-lesioned mouse striatum. The ratio of the number of surviving TH-positive cells to the total number of grafted cells was highest when ES cells were treated with SDIA for 12 days before transplantation. This ratio revealed that grafting cell colonies was more efficient for obtaining TH-positive cells in vivo than grafting cell suspensions. When we grafted a cell suspension of 2 x 10(5), 2 x 10(4), or 2 x 10(3) cells into the 6-OHDA-lesioned mouse striatum, we observed only a few surviving TH-positive cells. In conclusion, inducing DA neurons from mouse ES cells by SDIA for 12 days and grafting cell colonies into mouse striatum was the most effective method for the survival of TH-positive neurons in vivo. (C) 2002 Wiley-Liss, Inc.