Human embryonic² stem cells（胚胎干细胞） derived³ from excess IVF embryos⁴（晶胚） may help scientists unlock the mysteries of infertility⁵（不肥沃，不毛） for other couples struggling to conceive（构思，怀孕）, according to new research from the Stanford University School of Medicine. Researchers at the school have devised a way to efficiently⁶ coax⁷（哄，诱骗） the cells to become human germ cells — the precursors⁸ of egg and sperm⁹ cells — in the laboratory. Unlike previous research, which yielded primarily immature¹⁰ germ cells, the cells in this most-recent study functioned well enough to generate sperm cells. "Ten to 15 percent of couples are infertile," said senior author Renee Reijo Pera, PhD. "About half of these cases are due to an inability to make eggs or sperm. And yet deleting or increasing the expression of genes¹¹ in the womb to understand why is both impossible and unethical. Figuring out the genetic¹² 'recipe（处方，秘诀）' needed to develop human germ cells in the laboratory will give us the tools we need to trace what's going wrong for these people." Reijo Pera is a professor of obstetrics and gynecology（妇产科） at the medical school and the director of Stanford's Center for Human Embryonic Stem Cell Research and Education. The study will be published online by Nature on Oct. 28.

Previous efforts to study infertility have been hampered¹³ by the fact that — unlike many other biological processes — the human reproductive cycle cannot be adequately studied in animal models. And because germ cells begin to form very early in embryonic development (by eight to 10 weeks), there's been a dearth¹⁴（缺乏） of human material to work with. "Humans have a unique reproductive system," Reijo Pera said. "Until now we've relied on studies in mice to understand human germ cell differentiation¹⁵, but the reproductive genes are not the same. This is the first evidence that you can create functional¹⁶ human germ cells in a laboratory."

The scientists built on previous research in the mid-1990s by Reijo Pera that identified a number of genes involved in male infertility. Members of what's called the DAZ family, the genes are unusual in that they encode RNA-binding proteins rather than the DNA¹⁷ transcription factors more commonly known to regulate cellular¹⁸ events.

In the current study, the researchers treated human embryonic stem cells with proteins known to stimulate¹⁹ germ cell formation and isolated²⁰ those that began to express germ-cell-specific genes — about 5 percent of the total. In addition to expressing key genes, these cells also began to remove modifications²¹, or methyl groups（甲基原子团）, to their DNA that confer cell-specific traits that would interfere²² with their ability to function as germ cells. Such epigenetic（外成的，后生的） reprogramming is a hallmark of germ cell formation.

They then used a technique called RNA silencing to examine how blocking the expression of each of three DAZ family members in the embryonic stem cells affected²³ germ cell development. Conversely, they also investigated what happened when these genes were overexpressed.

They found that one family member, DAZL, functions very early in germ cell development, while two others, DAZ1 and BOULE, stimulate the then-mature germ cells to divide to form gametes（配子）. Overexpressing the three proteins together allowed the researchers to generate haploid cells（单倍体细胞） — those with only one copy of each chromosome²⁴（染色体） — expressing proteins found in mature sperm. (When a sperm and an egg join, the resulting fertilized²⁵ egg again has two copies of each chromosome.) When treated in this manner, about 2 percent of the differentiated²⁶ human embryonic stem cells were haploid（单一的） after 14 days of differentiation.

The effect of the DAZ family members on the embryonic stem cells varied²⁷ according to whether the cells were derived from a male or a female embryo¹. Overexpression of BOULE increased the relative proportion of putative²⁸ germ cells from 2 to 12 percent in female, but not male, cell lines. This suggests that BOULE may play a larger role than the other proteins in the development of female germ cells.

The researchers plan to use a similar strategy to optimize²⁹ the production of eggs from embryonic stem cells, as well as investigating whether reprogrammed adult cells called induced pluripotent（多能的） cells, or iPS cells, can also be used to create germ cells. By charting the milestones³⁰ of gamete development, they hope to identify potential problems that would lead to infertility or fetal disability.

"Although most of our birth defects are caused by problems in the development of eggs or sperm," said Reijo Pera, "it's not clear why there are so many errors. This research gives us a system we can use to compare errors in the germ line vs. somatic cells（体细胞）. For instance, we can now begin to directly investigate the effects of environmental toxins³¹（毒素） on germ cell differentiation and gamete development. We've already seen that, even in a dish, germ cells appear to be more sensitive to these compounds."