Why do cultured transplanted myoblasts die in vivo?: DNA quantification shows enhanced survival of donor male myoblasts in host mice depleted of CD4+ and CD8+ cells or NK1.1+ cells
Why do cultured transplanted myoblasts die in vivo?: DNA quantification shows enhanced survival of donor male myoblasts in host mice depleted of CD4+ and CD8+ cells or NK1.1+ cells
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DOI：
10.1177/096368970000900406
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发表时间：
2000-07-01
影响因子：
3.3
通讯作者：
Grounds, MD
中科院分区：
文献类型：
--
作者：
Hodgetts, SI;Beilharz, MW;Grounds, MD
克服注射的供体成肌细胞的大量和快速死亡是成肌细胞转移疗法（MTT）成功的主要障碍，该疗法被设计用于治疗致命的儿童肌病杜氏肌营养不良症。将雄性成肌细胞注射到雌性宿主小鼠中，并使用Y染色体特异性（Y1）探针定量存活的雄性DNA，可以确定供体成肌细胞死亡的速度和程度。将培养的正常C57 BL/10 Sn雄性供体成肌细胞注射到未处理的正常C57 BL/10 Sn和营养不良mdx雌性宿主小鼠中，并使用P-32标记的Y1探针通过狭缝印迹进行分析。来自供体成肌细胞的雄性DNA的量在几分钟内显示出显著减少，到1小时时仅占最初注射的2.5 × 10（5）个细胞的约10-18%（指定为100%）。这一数字在一周内进一步下降至约1- 4%。宿主环境（正常或营养不良）以及组织培养中的传代程度（早期“P3”或晚期“P15-20”传代）对此结果无差异。在注射培养的成肌细胞之前，通过CD 4（+）/CD 8（+）消耗抗体调节宿主反应，显著提高了营养不良mdx宿主中供体成肌细胞的存活率（1周后，相对于未处理的宿主为15倍）。与未处理的宿主相比，NK1.1缺失也显著提高了营养不良mdx宿主中供体成肌细胞的存活率（1周后21倍）。这些结果提供了一个战略性的方法，以提高供体成肌细胞的生存在MTT的临床试验。
Overcoming the massive and rapid death of injected donor myoblasts is the primary hurdle for successful myoblast transfer therapy (MTT), designed as a treatment for the lethal childhood myopathy Duchenne muscular dystrophy. The injection of male myoblasts into female host mice and quantification of surviving male DNA using the Y-chromosome-specific (Y1) probe allows the speed and extent of death of donor myoblasts to be determined. Cultured normal C57BL/10Sn male donor myoblasts were injected into untreated normal C57BL/10Sn and dystrophic mdx female host mice and analyzed by slot blots using a P-32-labeled Y1 probe. The amount of male DNA from donor myoblasts showed a remarkable decrease within minutes and by 1 h represented only about 10-18% of the 2.5 x 10(5) cells originally injected (designated 100%). This declined further over 1 week to approximately 1-4%. The host environment (normal or dystrophic) as well as the extent of passaging in tissue culture (early "P3" or late "P15-20" passage) made no difference to this result. Modulation of the host response by CD4(+)/CD8(+)-depleting antibodies administered prior to injection of the cultured myoblasts dramatically enhanced donor myoblast survival in dystrophic mdx hosts (15-fold relative to untreated hosts after 1 week). NK1.1 depletion also dramatically enhanced donor myoblast survival in dystrophic mdx hosts (21-fold after 1 week) compared to untreated hosts. These results provide a strategic approach to enhance donor myoblast survival in clinical trials of MTT.