Institute of Reconstructive Neurobiology, Life & Brain Center, University of Bonn and Hertie Foundation, Sigmund-Freud-Str. 25, 53105, Bonn, Germany.
Stem Cell Rev Rep. 2010 Jun;6(2):260-9. doi: 10.1007/s12015-010-9114-9.
Due to their pluripotency and their self-renewal capacity, human pluripotent stem cells (hPSC) provide fascinating perspectives for biomedical applications. In the long term, hPSC-derived tissue-specific cells will constitute an important source for cell replacement therapies in non-regenerative organs. These therapeutic approaches, however, will critically depend on the purity of the in vitro differentiated cell populations. In particular, remaining undifferentiated hPSC in a transplant can induce teratoma formation. In order to address this challenge, we have developed a laser-based method for the ablation of hPSC from differentiating cell cultures. Specific antibodies were directed against the hPSC surface markers tumor related antigen (Tra)-1-60 and Tra-1-81. These antibodies, in turn, were targeted with nanogold particles. Subsequent laser exposure resulted in a 98,9 +/- 0,9% elimination of hPSCs within undifferentiated cell cultures. In order to study potential side effects of laser ablation on cells negative for Tra-1-60 and Tra-1-81, hPSC were mixed with GFP-positive hPSC-derived neural precursors (hESCNP) prior to ablation. These studies showed efficient elimination of hPSC while co-treated hESCNP maintained their normal proliferation and differentiation potential. In vivo transplantation of treated and untreated mixed hPSC/hESCNP cultures revealed that laser ablation can dramatically reduce the risk of teratoma formation. Laser-assisted photothermolysis thus represents a novel contact-free method for the efficient elimination of hPSC from in vitro differentiated hPSC-derived somatic cell populations.
由于其多能性和自我更新能力，人类多能干细胞 (hPSC) 为生物医学应用提供了令人着迷的前景。从长远来看，hPSC 衍生的组织特异性细胞将成为非再生器官细胞替代疗法的重要来源。然而，这些治疗方法将严重依赖于体外分化细胞群体的纯度。特别是，移植中残留的未分化 hPSC 可诱导畸胎瘤形成。为了解决这一挑战，我们开发了一种基于激光的方法，用于从分化细胞培养物中消融 hPSC。特异性抗体针对 hPSC 表面标志物肿瘤相关抗原 (Tra)-1-60 和 Tra-1-81。这些抗体反过来又被纳米金颗粒靶向。随后的激光照射导致未分化细胞培养物中 98.9 +/- 0.9%的 hPSC 被消除。为了研究激光消融对 Tra-1-60 和 Tra-1-81 阴性细胞的潜在副作用，在消融之前，hPSC 与 GFP 阳性的 hPSC 衍生神经前体细胞 (hESCNP) 混合。这些研究表明，激光消融可以有效消除 hPSC，而共处理的 hESCNP 保持其正常的增殖和分化潜力。经处理和未经处理的混合 hPSC/hESCNP 培养物的体内移植表明，激光消融可以显著降低畸胎瘤形成的风险。因此，激光辅助光热解代表了一种从体外分化的 hPSC 衍生体细胞群体中有效消除 hPSC 的新型非接触方法。
