Division of Gynecologic Oncology, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, 75 Francis St., Boston, MA 02115, USA.
Division of Reproductive Endocrinology and Infertility, Center for Infertility and Reproductive Surgery, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St., Boston, MA 02115, USA.
EBioMedicine. 2023 Aug;94:104715. doi: 10.1016/j.ebiom.2023.104715. Epub 2023 Jul 21.
Treatment options for premature ovarian insufficiency (POI) are limited to hormone replacement and donor oocytes. A novel induced pluripotent stem cell (iPSC) transplant paradigm in a mouse model has potential translational applications for management of POI.
Mouse ovarian granulosa cell derived-iPSCS were labelled with green fluorescent protein (GFP) reporter and differentiated in vitro into oocytes. Differentiated cells were assayed for estradiol and progesterone secretion by enzyme-linked immunosorbent assays. After Fluorescence-Activated Cell Sorting (FACS) for the cell surface marker anti-Mullerian hormone receptor (AMHR2), enriched populations of differentiated cells were surgically transplanted into ovaries of mice that had POI secondary to gonadotoxic pre-treatment with alkylating agents. A total of 100 mice were used in these studies in five separate experiments with 56 animals receiving orthotopic ovarian injections of either FACS sorted or unsorted differentiated iPSCSs and the remaining animals receiving sham injections of PBS diluent. Following transplantation surgery, mice were stimulated with gonadotropins inducing oocyte development and underwent oocyte retrieval. Nine transplanted mice were cross bred with wild-type mice to assess fertility. Lineage tracing of resultant oocytes, F1 (30 pups), and F2 (42 pups) litters was interrogated by GFP expression and validation by short tandem repeat (STR) lineage tracing.
[1] iPSCs differentiate into functional oocytes and steroidogenic ovarian cells which [2] express an ovarian (GJA1) and germ cell (ZP1) markers. [3] Endocrine function and fertility were restored in mice pretreated with gonadotoxic alkylating agents via orthotopic transplantation of differentiated iPSCS, thus generating viable, fertile mouse pups.
iPSC-derived ovarian tissue can reverse endocrine and reproductive sequelae of POI.
Center for Infertility and Reproductive Surgery Research Award, Siezen Foundation award (RMA). Reproductive Scientist Development Program, Marriott Foundation, Saltonstall Foundation, Brigham Ovarian Cancer Research Fund (K.E).
治疗卵巢早衰（POI）的方法有限，仅限于激素替代和供体卵。一种新型的诱导多能干细胞（iPSC）移植范式在小鼠模型中具有管理 POI 的潜在转化应用。
用绿色荧光蛋白（GFP）报告基因标记小鼠卵巢颗粒细胞来源的 iPSCs，并在体外分化为卵母细胞。通过酶联免疫吸附试验检测分化细胞的雌二醇和孕酮分泌。荧光激活细胞分选（FACS）后，用抗苗勒管激素受体（AMHR2）的细胞表面标志物对分化细胞进行富集，然后将富集的分化细胞群体通过手术移植到因烷化剂性腺毒性预处理而发生 POI 的小鼠卵巢中。在五项独立实验中，共有 100 只小鼠参与了这些研究，其中 56 只动物接受了 FACS 分选或未分选的分化 iPSCS 的卵巢原位注射，其余动物接受了 PBS 稀释剂的假注射。移植手术后，用促性腺激素刺激小鼠诱导卵母细胞发育，并进行卵母细胞回收。将 9 只移植小鼠与野生型小鼠杂交，以评估其生育能力。通过 GFP 表达和短串联重复（STR）谱系追踪验证，对所得卵母细胞、F1（30 只幼崽）和 F2（42 只幼崽）后代的谱系追踪进行了检测。
[1] iPSCs 分化为功能性卵母细胞和类固醇生成性卵巢细胞，[2] 表达卵巢（GJA1）和生殖细胞（ZP1）标志物。[3] 通过将分化的 iPSCS 原位移植到接受性腺毒性烷化剂预处理的小鼠中，恢复了内分泌和生殖功能，从而产生了有活力的、可育的小鼠幼崽。
iPSC 衍生的卵巢组织可以逆转 POI 的内分泌和生殖后遗症。
生育和生殖外科研究奖中心，Siezen 基金会奖（RMA）。生殖科学家发展计划、万豪基金会、Saltonstall 基金会、布莱根卵巢癌研究基金（K.E.）。
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