Ultrastructural characteristics of human granulosa cells in a coculture system for in vitro fertilization
Ultrastructural characteristics of human granulosa cells in a coculture system for in vitro fertilization
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体外受精共培养系统中人颗粒细胞的超微结构特征
DOI：
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
G. Macchiarelli
中科院分区：
文献类型：
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作者：
S. Nottola;R. Heyn;Alessandra Camboni;S. Correr;G. Macchiarelli
体外受精（IVF）过程中体细胞共培养的使用目前已经完成，以获得更多的健康和有活力的胚胎，具有较高的植入潜力。在可用作共培养饲养细胞的不同细胞系中，颗粒细胞（GC）是安全且易于回收的自体细胞。本研究的目的是分析用于共培养系统中的人GCs的精细结构，以从形态动力学的角度评估它们在支持胚胎发育中的作用。在人绒毛膜促性腺激素给药后36小时，从接受IVF手术的患者中采集卵母细胞，这些患者已提供知情同意书以纳入本方案。共培养后，固定GC并进行光学显微镜（LM）和透射电子显微镜（TEM）检查。光镜下，GCs呈不规则圆形或多面形，细胞排列疏松，直径18 ~ 25 μm。偶尔观察到有丝分裂细胞以及退化元件（具有固缩核或致密细胞质）和细胞碎片。透射电镜观察，由于胞质外翻，质膜呈不规则形。相邻的GC之间的线性和环形缝隙连接被发现。GC核，圆形和偏心定位，含有精细分散的染色质，一个（通常是两个）突出的核仁，很少，异染色质的周边补丁。GC细胞质中有大量细胞器，其中线粒体呈杆状或细长形，通常具有管状泡状嵴，但偶尔显示非典型的纵向嵴。在GC细胞质中还观察到滑面内质网膜、高尔基体和囊泡、分泌样颗粒、粗面内质网（RER）池、游离核糖体和多聚核糖体、溶酶体样小体、微丝和脂滴。大多数细胞的粗面内质网几乎不存在，核周间隙充满大量脂滴。相反，一些GC含有丰富的粗面内质网和罕见的脂滴分散在细胞质中。总之，我们的数据表明，在共培养系统中，GC的存在提供了典型的健康，代谢活性，主要是类固醇生成细胞的超微结构特征。还发现了蛋白质合成细胞。根据生物化学和临床研究评估的这些数据，维持了人GC共培养物通过分泌类固醇和蛋白质（推定的“胚胎营养”因子）对体外早期胚胎发育产生积极影响的能力。Microsc. Res. Tech.，2006.© 2006 Wiley利斯公司
The use of somatic cells for cocultures during in vitro fertilization (IVF) is currently finalized to obtain a higher number of healthy and viable embryos with a high potential of implantation. Among the different cell lines that can be used as feeder cells for cocultures, granulosa cells (GCs) are autologous cells, safe and easy to recover. The aim of the present study was to analyze the fine structure of human GCs used in a coculture system to evaluate, from a morphodynamic point of view, their role in supporting embryo development. GCs were collected during oocyte pick‐up, 36 h after human chorionic gonadotropin administration, from patients undergoing IVF procedures, who had given their informed consent to be included in this protocol. After coculture, GCs were fixed and processed for light microscopy (LM) and transmission electron microscopy (TEM). By LM, GCs appeared as clusters of loosely packed cells, irregularly rounded or polyhedral in shape, varying in diameter from 18 to 25 μm. Mitotic cells, as well as regressing elements (with pyknotic nuclei or dense cytoplasm) and cell fragments were occasionally observed. By TEM, the plasma membrane was irregular due to the presence of cytoplasmic evaginations. Linear and annular gap junctions between neighboring GCs were found. GC nuclei, rounded and eccentrically located, contained finely dispersed chromatin, one (often two) prominent nucleoli and, infrequently, peripheral patches of heterochromatin. Numerous organelles populated the GC cytoplasm, among them, mitochondria were rod‐shaped or elongated, usually provided with tubular–vesicular cristae but occasionally showing atypical, longitudinally oriented cristae. Membranes of smooth endoplasmic reticulum, Golgi stacks and vesicles, secretory‐like granules, cisternae of rough endoplasmic reticulum (RER), free ribosomes and polysomes, lysosomal‐like bodies, microfilaments, and lipid droplets were also seen in the GC cytoplasm. In most cells, RER was scarcely represented and numerous lipid droplets filled the perinuclear space. On the contrary, some GCs contained an abundant RER and rare lipid droplets scattered in the cytoplasm. In conclusion, our data demonstrated the presence, in a coculture system, of GCs provided with ultrastructural characteristics typical of healthy, metabolically active, mostly steroidogenic cells. Protein‐synthetic cells have also been detected. These data, evaluated at the light of biochemical and clinical studies, sustain the capability of human GCs cocultures to positively affect early embryo development in vitro by the secretion of steroids and proteins, putative “embryotrophic” factors. Microsc. Res. Tech., 2006. © 2006 Wiley‐Liss, Inc.
胶原基质影响人颗粒细胞的形态特征和类固醇分泌。
影响因子：
9.8
作者：
Ben-Rafael,Z;Benadiva,CA;MastroianniJr,L;Garcia,CJ;Minda,JM;Iozzo,RV;Flickinger,GL
通讯作者：
Flickinger,GL
促性腺激素和环腺苷 3,5-单磷酸 (cAMP) 会改变培养的人颗粒细胞的形态。
影响因子：
3.6
作者：
Soto,EA;Kliman,HJ;Strauss3rd,JF;Paavola,LG
通讯作者：
Paavola,LG