Ceska Gynekol. 2021;86(1):5-10. doi: 10.48095/cccg20215.
The work deals with a clinical part of human embryonic stem cell (hESC) research. The aim of the project is the differentiation of somatic cell types, useful in drug development, regenerative medicine and cell therapy. The aim of this work is to enable targeted therapy of yet incurable diseases. The pluripotent hESCs have unlimited self-renewal capacity. This ability is used in therapy to create missing or damaged cells in the human body. It is of interest to develop clinical-grade hESC lines useful in preclinical and clinical studies.
The derivation of the hESC must respect the legislation of the Czech Republic and the EU. The aim was to develop an informed consent of both donors for donated discarded embryos that are not suitable for treatment by in vitro fertilization according to Directive 2004/23/EC. The FNBs Center for Assisted Reproduction (CAR) participates in oocyte collection, cultivation and cryopreservation of embryos, communication with clients and ensuring the informed consent of embryo donors. A transport protocol and a methodology for handing over the thawed embryos with the original numerical code were developed. Before the embryos are handed over to the ICRC co-authors workplace (CTEF), they are thawed and, if necessary, recultivated to the blastocyst stage; afterwards, assisted hatching is performed.
In the period from January 2018 to July 2020, 138 selected suitable clients were asked for donations, with 52 not responding, 19 terminating and 29 extending the embryo storage. Only 38 clients, i.e. 27.5%, agreed with the usage of their embryos for the preparation of hESCs. In the same period, personal communication with suitable CAR clients took place and another 17 embryo donors were obtained. A total of 160 embryos were obtained from 55 donors aged 26 to 42 years. The embryos were most often frozen in the blastocyst (53 embryos - 33.1%) and morula (74 embryos - 46.3%) stages. Of the 29 genetically examined embryos, only 5 are euploid (17.2%), 2 are mosaic and 22 are aneuploid or with translocations or carriers with a monogenic defect.
We have an informed consent prepared and approved by the Ethics Committee of the Masaryk University and the University Hospital Brno; 160 donated embryos have been selected and secured. A transport protocol and handover methodology are developed. The plan for the transfer of thawed anonymized embryos in the first phase, October - December 2020, includes approximately 5 thawed blastocysts per week with assisted hatching. After their transfer to the CTEF, the embryoblast will be isolated with subsequent cultivation. The established hESCs must meet the specified criteria of safety, stability and pluripotency. We believe that, in accordance with the project plan, we will obtain at least 3 clinical-grade hESC lines, the first created in the Czech Republic, respecting the requirements for Advanced Medicinal Therapy Products (AMTP).
本工作涉及人类胚胎干细胞（hESC）研究的临床部分。该项目的目的是分化体细胞类型，这些类型在药物开发、再生医学和细胞治疗中有用。本工作的目的是实现对目前仍无法治愈的疾病的靶向治疗。多能 hESC 具有无限的自我更新能力。在治疗中，这种能力被用于在人体内创造缺失或受损的细胞。开发可用于临床前和临床研究的临床级 hESC 系是很有意义的。
hESC 的衍生必须遵守捷克共和国和欧盟的立法。本研究旨在根据指令 2004/23/EC 为不适合体外受精治疗的废弃胚胎的捐献者制定知情同意书。辅助生殖（CAR）的 FNBs 中心参与卵母细胞的收集、培养和胚胎的冷冻保存、与客户的沟通以及确保胚胎捐献者的知情同意。制定了解冻胚胎的运输方案和原始编号移交方法。在将解冻的胚胎移交给 ICRC 合著者工作场所（CTEF）之前，对其进行解冻，如果需要，将其培养到囊胚阶段；之后，进行辅助孵化。
在 2018 年 1 月至 2020 年 7 月期间，向 138 名合适的客户询问了捐赠事宜，其中 52 名未回复，19 名终止，29 名延长了胚胎储存时间。只有 38 名客户（27.5%）同意将其胚胎用于制备 hESC。在同一时期，与合适的 CAR 客户进行了个人沟通，并获得了另外 17 名胚胎供体。总共从 55 名 26 至 42 岁的供体中获得了 160 个胚胎。胚胎最常处于囊胚（53 个-33.1%）和桑椹胚（74 个-46.3%）阶段冷冻。在 29 个经过基因检测的胚胎中，只有 5 个是整倍体（17.2%），2 个是嵌合体，22 个是非整倍体或带有易位或携带单基因缺陷。
我们已经准备并获得了马萨里克大学和布尔诺大学附属医院伦理委员会的知情同意书；已经选择并保护了 160 个捐赠的胚胎。制定了运输方案和移交方法。2020 年 10 月至 12 月第一阶段解冻匿名胚胎的转移计划包括每周大约 5 个辅助孵化的解冻囊胚。在将其转移到 CTEF 后，将分离胚胎原代细胞并随后进行培养。建立的 hESC 必须符合安全性、稳定性和多能性的规定标准。我们相信，按照项目计划，我们将获得至少 3 种临床级 hESC 系，这是在捷克共和国首次创建的，符合高级治疗药物产品（AMTP）的要求。
