HLA-Matching Potential of an Established Human Embryonic Stem Cell Bank in China
HLA-Matching Potential of an Established Human Embryonic Stem Cell Bank in China
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中国已建立的人胚胎干细胞库的HLA匹配潜力
DOI：
10.1016/j.stem.2009.10.009
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发表时间：
2009-11-06
期刊：
影响因子：
23.9
通讯作者：
Lu, Guangxiu
中科院分区：
文献类型：
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作者：
Lin, Ge;Xie, Yubin;Lu, Guangxiu
先前的研究已经对来自任一尸体器官供体的HLA和ABO血型数据进行了内部匹配（Taylor等人，2005）或脐带血库（Nakajima等人，2007）来估计具有各种HLA基因型的约150-170个人胚胎干细胞（hESC）系的理论集合或对于常见HLA类型纯合的10个系的集合将足以为英国的群体提供HLA匹配（Taylor等人，2005）和日本（Nakajima等人，2007年）。在这份报告中，我们已经建立了一个hESC银行包含188 hESC线主要来自异常和低质量的体外受精（IVF）衍生的胚胎，并评估了实际的HLA匹配能力的银行相对于区域人口。我们发现，目前的银行可以提供一个有益的匹配24.9%-56.3%的当地人口，无论种族差异。尽管hESC对免疫反应较不敏感（Drukker等人，2006），这些细胞及其衍生物可以表达人白细胞抗原（HLA），并且随后的主要组织相容性复合体（MHC）不相容性可以在移植后引起同种异体移植物排斥（布拉德利等人，2002年; Drukker和Benvenisty，2004年）。以前的理论计算在实际情况中没有阐明（1）来自体外受精（IVF）治疗的胚胎是否可以提供足够的HLA多样性用于匹配;（2）Taylor等和Nakajima等研究中用于建模计算的假定群体是否准确地反映了现有群体;以及（3）不同地理区域中存在的种族组成的多样性是否影响HLA配型所需的hESC系的数量。此外，破坏大量正常人类胚胎以获得细胞系是一个伦理上有争议的问题;因此，我们进行了这项研究，以评估从不适合IVF胚胎移植（ET）的胚胎中分离的细胞系是否可以为库细胞系来源的区域人群提供合适的HLA匹配。在获得两个配子供体的书面知情同意书和医院伦理委员会的批准后，从各种异常胚胎中共收集了692个囊胚。收集的囊胚不适合IVF-ET，因为它们是从受精后第3天的低质量胚胎中获得的，在受精卵阶段具有异常数量的原核，在植入前遗传学诊断（PGD）中显示出异常，或从人工孤雌激活的卵母细胞中获得。采用无血清机械衍生方法以促进这些细胞系的潜在临床应用（参见在线提供的补充实验程序），并成功建立了188个hESC系。我们观察到单个囊胚的内细胞团（ICM）质量与衍生效率相关。当我们使用具有高质量或中等质量ICM（A级和B级，如前所述[Gardner等人，2000年）。在我们的研究中，69个细胞系来自170个具有A级ICM的囊胚（40.6%）; 99个细胞系来自332个具有B级ICM的囊胚（29.8%）; 6个细胞系来自83个具有C级ICM的囊胚（7.2%）（表1和图S1）。ICM等级反映了ICM的大小和紧凑程度;因此，高等级的ICM可以产生更多的多能细胞，从而有利于更有效的衍生。影响衍生效率的另一个因素是ICM的发展阶段。
Previous studies have performed internal matching of HLA and ABO blood type data from either cadaveric organ donors (Taylor et al., 2005) or a cord blood bank (Nakajima et al., 2007) to estimate that a theoretical collection of approximately 150–170 human embryonic stem cell (hESC) lines with various HLA genotypes or a collection of ten lines homozygous for the common HLA types will suffice to provide HLA-matches for populations in the UK (Taylor et al., 2005) and Japan (Nakajima et al., 2007). In this report, we have established a hESC bank containing 188 hESC lines mainly derived from abnormal and poor-quality in vitro fertilization (IVF)-derived embryos and have evaluated the actual HLA-matching capacity of the bank with respect to the regional population. We find that the current bank could provide a beneficial match for 24.9%–56.3% of the local population, regardless of ethnic differences. Although hESCs are less susceptible to immune reaction (Drukker et al., 2006), these cells and their derivatives can express human leukocyte antigen (HLA), and the consequent major histocompatibility complex (MHC) incompatibility can cause allograft rejection after transplantation (Bradley et al., 2002; Drukker and Benvenisty, 2004). The previously theoretical calculations did not clarify in actual situation (1) whether the embryos from in vitro fertilization (IVF) treatment can provide adequate HLA diversity for matching;(2) whether the presumptive populations used for modeling calculations in the Taylor et al. and Nakajima et al. studies accurately reflect existing populations; and (3) whether diversity in the ethnic composition present in different geographical regions influences the number of hESC lines required for HLA matching. Moreover, the destruction of large numbers of normal human embryos to obtain cell lines is an ethically controversial issue; thus, we undertook this study to assess whether lines isolated from embryos unsuitable for IVF embryo transfer (ET) could offer a suitable HLA-match for the regional population from which the banked lines were derived. A total of 692 blastocysts obtained from various abnormal embryos were collected after obtaining written informed consent from both gamete donors and the approval of the hospital ethical committee. The collected blastocysts were not suitable for IVF-ET because they were obtained from poor-quality embryos on day 3 postfertilization, had an abnormal number of pronuclei at the zygote stage, showed abnormalities in the preimplantation genetic diagnosis (PGD), or were obtained from artificially parthenoactivated oocytes. A serum-free mechanical derivation method was adopted to facilitate the potential clinical application of these cell lines (see Supplemental Experimental Procedures available online), and 188 hESC lines were successfully established. We observed that the quality of the inner cell mass (ICM) of individual blastocysts correlated with the derivation efficiency. The rate of successful isolation of hESC lines was significantly higher when we used blastocysts with high-or medium-quality ICMs (grade A and B, as previously described [Gardner et al., 2000]). In our study, 69 cell lines were derived from 170 blastocysts with grade A ICMs (40.6%); 99 cell lines were derived from 332 blastocysts with grade B ICMs (29.8%); and 6 cell lines were derived from 83 blastocysts with grade C ICMs (7.2%)(Table 1 and Figure S1). The ICM grade reflects the size and degree of compactness of ICM; therefore, highgrade ICMs may yield more pluripotent cells and thus favor more efficient derivation. Another factor affecting the derivation efficiency is the developmental stage of the ICM used for …