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人类卵细胞实验室首度成长成功

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科学家们已首度於实验室成功将人类未成熟卵细胞成长为成熟卵细胞。这项技术,最终可能帮助拯救不符合传统卵细胞孕育女性癌症病人生育。

西北大学的研究人员自接受化疗前希望保留生育的14名妇女取用未成熟卵细胞,装在一个称为卵泡的保护囊,将细胞置於独特的三维生长环境30天,耐心处理细胞,将其成为健康的,具功能的人类卵细胞。

“这是首度突破”,圣路易斯圣卢克医院没有参与这项研究的不孕专家西尔伯谢尔曼说。“还没有人测试了这些卵细胞的体外受精和怀孕,但他们看上去很正常,我们都感到兴奋。”

传统维护女性癌症患者的生育能力方式是自其卵巢以手术移除完全成熟卵子,在实验室中立即受精,并冻结所成的胚胎。但是由于每月排卵只有一个卵泡成熟,这种方法需要2到6周的激素治疗,以产生足够的成熟卵子来做受精。

“癌症患者通常没有这么多时间浪费,”西尔伯说。“并且要给她任何形式的保证,便不得不把她通过三至六个月或更多这样的周期,以获得足够的适量的卵子。冷冻卵巢组织,我们将获得数以万计的卵子数,使病人能得到更大程度的安全感以应其将来的生育。“

此外,高剂量的激素对某些类型癌症的患者是很危险的,如乳腺癌或卵巢肿瘤,而且传统的方法是不适於没有经历过青春期的女孩。如果医生可以采取未成熟卵泡,体外成长成卵子,激素使用便可完全避免。

但至今,没有人能在实验室成长卵泡。先前大多都尝试涉及二维环境的细胞培养,但任何形式的压力便会抑制卵细胞的生长。

“科学家们多年来使用一块平面塑料”,20日刊登在人类生殖论文的共同作者西北大学生育研究员德丽莎伍德拉夫说。“当如此作,周围的细胞会开始移走,和卵子之间的联系及支持细胞便会遗失。”

支持细胞很是关键,伍德拉夫说,因为他们提供卵子成长需求的激素和营养物。要创造卵泡理想的生长环境,研究人员便与精通生物材料的生物医学工程师合作。

“我们的突破是使用所谓的海藻酸钠水凝胶,不会触及或联系卵泡,而只会支持它们”,伍德拉夫说。原来僵化的凝胶对卵泡功能至关重要的:如果凝胶过于僵硬,卵泡发病而产生错误的荷尔蒙。“我们很幸运自一开始就使用了非常柔软的凝胶”,她说。

在三维矩阵卵泡孵化后的30天,研究人员发现,它们已经成长为成熟卵子的规模,而且产生正确和正确数量的荷尔蒙,如雌激素和黄体酮。

真正考验实验室生长的卵子是要观查是否可以进行最后阶段的细胞分裂及孕育。

“我们用在老鼠,自开始到孕育和活产幼鼠”,伍德拉夫说。但是,美国国立卫生研究院条例不允许科学家作人类卵子受精的研究 -- 最后一步的程续必须由不孕症诊所医生在自愿生育的病人身上完成。

“该证明是将卵子在体外受精的和孕育出婴儿”,西尔伯说。“但这是额外的阶段,这令人感到惊讶,他们已如此深入“。

“10年前,这被认为是不可能完成的任务,我们当时也认为再过50年才会发生 ”,他说。“令人吃惊的是,它比我们想像的简单得多”。
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译文:A Fertility First: Human Egg Cells Grow Up in Lab


For the first time, scientists have managed to grow mature humaneggs from immature cells in the lab, a technique that may eventuallyhelp save the fertility of female cancer patients who aren’t eligiblefor traditional egg harvest.

Researchers from Northwestern University took immature egg cells,encased in a protective sac called a follicle, from 14 women who wantedto preserve their fertility before undergoing chemotherapy. By placingthe cells in a unique three-dimensional growing environment for 30days, the scientists coaxed the cells into becoming what appear to behealthy, functional human eggs.

“It is a major first,” said infertility expert Sherman Silber of St.Luke’s Hospital in St. Louis, who was not involved in the research. “Noone has yet tested the eggs by in-vitro fertilization and pregnancy,but they look quite normal and we are all excited about it.”

The traditional way to preserve a female cancer patient’s fertilityis to surgically remove fully mature eggs from her ovary, fertilize theeggs immediately in the lab, and freeze the resulting embryos. Butsince only one follicle matures each month for ovulation, that methodrequires two to six weeks of hormone therapy to generate enough matureeggs for harvest.

“The cancer patient usually does not have this much time to waste,”Silber said. “And to give her any kind of assurance, you would have toput her through three to six or more such cycles to get enough eggs tobe comfortable. With ovary tissue freezing, we will get hundreds ofthousands of eggs, and the patient can get a much greater measure ofsecurity for her future fertility.”

In addition, giving high doses of hormones is dangerous for patientswith certain types of cancer, such as breast or ovarian tumors, and thetraditional method won’t work for girls who haven’t gone throughpuberty. If physicians could take immature ovarian follicles and growthem into eggs outside the body, they could skip the hormone stepaltogether.

Until now, however, no one has been able to grow human ovarianfollicles in the lab. Most previous attempts had involved trying toculture the cells in a two-dimensional environment, but it turns outthat any kind of pressure on the egg cells inhibits their growth.

“Scientists had been putting them on a flat piece of plastic foryears,” said fertility researcher Teresa Woodruff of NorthwesternUniversity, a co-author on the paper published Monday in Human Reproduction.“When you do that, the cells around the egg begin to move away, and theconnection between the egg and its supporting cells is lost.”

The supporting cells are critical, Woodruff said, because theyprovide the hormones and nutrients that the egg needs to grow. Tocreate the ideal growing environment for a follicle, the researcherscollaborated with biomedical engineers who specialize in biomaterials.

“Our breakthrough was to use a hydrogel called alginate, whichdoesn’t touch or contact the follicle cells, but just supports them,”Woodruff said. It turns out that the rigidity of the gel is crucial tofollicle function: If the gel is too stiff, the follicles start lookingsick and making the wrong kind of hormones. “We kind of lucked out inthe very beginning in that we used a very soft gel,” she said.

After incubating the follicles for 30 days in the three-dimensionalmatrix, the researchers discovered that they had grown to the size ofmature eggs and were producing all the right hormones, such as estrogenand progesterone, in the right quantities.

The true test of the lab-grown eggs will be to see if they canundergo a final stage of cell division to be ready for fertilization.

“We did it in a mouse and got it all the way to fertilization andlive birth,” Woodruff said. But regulations from the NationalInstitutes of Health won’t let scientists fertilize human eggs forresearch — the final step of the process must be done by physicians inan infertility clinic for a patient who’s ready to have a baby.

“The proof would be if they could fertilize the eggs in vitro andget babies out of it,” Silber said. “But that’s an extra stage, andit’s amazing they’ve got this far.”

“Ten years ago it was considered such an impossible task that wedidn’t think it would happen for another 50 years,” he said. “Theamazing thing is that it turned out to be much simpler than we everdreamed.”


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