Cellular transplantation for the nervous system: impact of time after preparation on cell viability and survival Laboratory investigation
Cellular transplantation for the nervous system: impact of time after preparation on cell viability and survival Laboratory investigation
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DOI：
10.3171/2009.10.jns09252
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发表时间：
2010-09-01
影响因子：
4.1
通讯作者：
Uram, Martin
中科院分区：
文献类型：
--
作者：
Gobbel, Glenn T.;Kondziolka, Douglas;Uram, Martin
目的。细胞移植已显示出治疗各种神经系统疾病的希望，但对移植后影响细胞存活和整合的因素知之甚少。事实上，人们对简单但潜在关键变量（包括细胞制备和给药方法）如何影响移植成功知之甚少。本研究的目的是确定组织制备和植入之间的时间对细胞活力的影响。时间可能因细胞准备、运送到手术室和手术技术而异。这项研究还旨在评估各种评估种植体活力的方法的敏感性。方法。神经祖细胞和骨髓基质细胞的细胞系是从健康成年小鼠中产生的。实验当天，收集细胞，悬浮在平衡盐溶液中，并根据相差显微镜下的外观、保留荧光染料的能力以及在 24 小时内与培养表面的附着情况，连续评估长达 3.5 小时的活力。结果。当根据细胞保留荧光染料的能力来测量活力时，活力每小时下降 10-15%。根据细胞附着在培养物表面并生长 24 小时的能力，活力下降得更快，每小时大约 20%。此外，基于相差显微镜或急性染料保留判断为存活的细胞中，只有约三分之一在平板接种中被发现是存活的，并且最初判断为存活的细胞中，只有 10% 在悬浮液中 3 小时后仍然能够存活。结论。作者的结果表明，在制备和植入之间，活力可能会显着降低，并且可能需要更复杂的评估方法，例如细胞附着到基质上并生长的能力，以检测活力的降低。准备和植入之间的时间将是临床试验设计的一个重要因素。 （DOI：10.3171/2009.10.JNS09252）
Object. Cell transplantation has shown promise for the treatment of various neurological disorders, but the factors that influence cell survival and integration following transplantation are poorly understood. In fact, little is known regarding how simple but potentially critical variables, including the method of cellular preparation and administration, might affect transplant success. The goal of the present study was to determine the impact of time between tissue preparation and implantation on cellular viability. Time can vary with cell preparation, delivery to the operating room, and surgical technique. This study was also designed to evaluate the sensitivity of various methods of assessing implant viability.Methods. Cell lines of neural progenitor cells and bone marrow stromal cells were generated from healthy adult mice. On the day of experimentation, the cells were collected, suspended in a balanced salt solution, and sequentially assessed for viability for up to 3.5 hours based on their appearance under phase-contrast microscopy, their ability to retain a fluorescent dye, and their attachment to a cultivation surface for 24 hours.Results. When viability was measured based on the ability of cells to retain a fluorescent dye, there was a decrease in viability of 10-15% each hour. Based on the ability of the cells to attach to a culture surface and grow for 24 hours, viability decreased more rapidly at approximately 20% per hour. In addition, only about one-third of the cells judged viable based on phase-contrast microscopy or acute dye retention were found to be viable based on plating, and only 10% of the cells initially judged as viable were still capable of survival after 3 hours in suspension.Conclusions. The authors' results indicate that that there can be significant losses in viability between preparation and implantation and that more sophisticated methods of evaluation, such as the ability of cells to attach to a substrate and grow, may be required to detect decreases in viability. The time between preparation and implantation will be an important factor in clinical trial design. (DOI: 10.3171/2009.10.JNS09252)