本文選題：卵母細(xì)胞 + 體外成熟　； 參考：《山東大學(xué)》2007年碩士論文

【摘要】： 研究目的：本研究旨在通過對人未成熟和體外培養(yǎng)成熟后卵母細(xì)胞中線粒體的分布的比較，闡明卵母細(xì)胞成熟和線粒體分布的關(guān)系。 研究方法：(1)未成熟卵母細(xì)胞的來源：未成熟卵母細(xì)胞來源于自2006年3月至10月就診于山東大學(xué)山東省立醫(yī)院生殖醫(yī)學(xué)中心，僅單純男方因素或同時(shí)合并女方輸卵管因素行卵母細(xì)胞內(nèi)單精子顯微注射的不孕癥患者。經(jīng)透明質(zhì)酸酶消化后，所獲未成熟卵母細(xì)胞經(jīng)患者同意，并簽署相關(guān)知情同意書后捐獻(xiàn)用于醫(yī)學(xué)科學(xué)研究。 (2)體內(nèi)成熟的卵母細(xì)胞的來源：體內(nèi)成熟的卵母細(xì)胞來源于同期在山東大學(xué)山東省立醫(yī)院生殖醫(yī)學(xué)中心就診，行促排卵后同房或人工授精的不孕癥患者，若注射HCG10 000IU 42 h后仍未排卵，則行人工破卵治療。即將大卵泡通過經(jīng)陰道穿刺的方法使之破裂，卵泡液流出，同時(shí)卵子也進(jìn)入腹腔，患者將有妊娠的可能。如果卵泡較多，為避免卵巢過度刺激綜合征和多胎妊娠，會(huì)對部分小卵泡進(jìn)行穿刺，穿刺獲得的成熟卵母細(xì)胞經(jīng)患者同意，并簽署相關(guān)知情同意書后捐獻(xiàn)用于醫(yī)學(xué)科學(xué)研究。 將未成熟卵母細(xì)胞隨機(jī)分為兩組，采用熒光染色法，一組直接染色，另一組體外培養(yǎng)成熟后再進(jìn)行染色。未成熟卵母細(xì)胞和成熟卵母細(xì)胞均用Mito Tracker Green FM染色，經(jīng)多聚甲醛固定后，，在激光掃描共聚焦顯微鏡下觀察線粒體的分布。 結(jié)果：284個(gè)未成熟卵母細(xì)胞中有144個(gè)進(jìn)行體外成熟培養(yǎng)，其中GV期卵母細(xì)胞88個(gè)，MI期卵母細(xì)胞56個(gè)。IVM 48h后，56個(gè)GV期卵母細(xì)胞達(dá)到MⅡ期，成熟率為63.64％；50個(gè)MⅠ期卵母細(xì)胞達(dá)到MⅡ期，成熟率為89.29％。MⅠ期卵母細(xì)胞的成熟率明顯高于GV期卵母細(xì)胞，兩者之間的差別有統(tǒng)計(jì)學(xué)意義(P=0.016)。 線粒體在細(xì)胞質(zhì)中的分布方式可分為以下三種：周邊分布、半周邊分布和均勻分布。在64.10％(50／78)的GV期卵母細(xì)胞中，線粒體呈周邊分布。在MⅠ期卵母細(xì)胞中，線粒體呈均勻分布的占38.71％(24／62)，45.16％(28／62)的MⅠ期卵母細(xì)胞的線粒體仍維持周邊分布。體外培養(yǎng)成熟以后，線粒體在75.47％(80／106)的卵母細(xì)胞中呈現(xiàn)均勻分布。GV期卵母細(xì)胞中線粒體以周邊分布為主，與MⅠ期和MⅡ期卵母細(xì)胞相比，其差異有統(tǒng)計(jì)學(xué)意義。成熟卵母細(xì)胞中線粒體大多呈均勻分布，與未成熟卵母細(xì)胞相比，也存在顯著差異。由此可以看出，人卵母細(xì)胞成熟前后，線粒體發(fā)生了明顯的分布變化，由未成熟卵母細(xì)胞中以周邊分布為主變?yōu)槌墒炻涯讣?xì)胞中以均勻分布為主。 體內(nèi)成熟的11個(gè)卵母細(xì)胞中，線粒體呈均勻分布的有8個(gè)(72.73％)，另有3個(gè)呈周邊分布(27.27％)。與體外培養(yǎng)成熟的卵母細(xì)胞相比，體內(nèi)成熟的卵母細(xì)胞中線粒體分布的特點(diǎn)是胞質(zhì)中央?yún)^(qū)域線粒體的濃集，熒光強(qiáng)度高于周邊區(qū)域。 結(jié)論：(1)人卵母細(xì)胞成熟前后，線粒體出現(xiàn)了明顯的分布變化，由未成熟卵母細(xì)胞中以周邊分布為主變?yōu)槌墒炻涯讣?xì)胞中以均勻分布為主。(2)體內(nèi)成熟的卵母細(xì)胞中線粒體的分布與體外成熟的卵母細(xì)胞略有不同。
[Abstract]:Objective: This study aims to elucidate the relationship between the maturation of oocyte and the distribution of mitochondria by comparing the distribution of mitochondria in the immature and in vitro cultured oocytes.
Research methods: (1) the source of immature oocyte: immature oocyte originates from March 2006 to October in the reproductive medical center of Shangdong Province-owned Hospital of Shandong University. Only male factor or single sperm microinjection in oocyte with female oviduct factors combined with female fallopian tube factors and hyaluronidase elimination. After that, the obtained oocytes were donated to medical research by consent of the patients and signing the relevant informed consent.
(2) the source of mature oocytes in the body: mature oocytes from the body of the body of the body from the same period at the reproductive medical center of Shangdong Province-owned Hospital of Shandong University, and the infertile patients with ovulatory or artificial insemination after ovulation induction. If HCG10 000IU 42 h were not ovulation, artificial ovulation was performed. The large follicles were punctured through the transvaginal puncture. The method makes it broken, the follicular fluid outflows and the egg enters the abdominal cavity. The patient will have the possibility of pregnancy. If there are more follicles, in order to avoid ovarian hyperstimulation syndrome and multiple pregnancy, some small follicles will be punctured, the mature oocyte obtained by the puncture is approved by the patient, and the related informed consent is signed and donated to medicine. Scientific research.
The immature oocytes were randomly divided into two groups, which were stained by fluorescent staining, a group of direct staining, and the other group was stained after maturation in vitro. The immature oocyte and mature oocyte were stained with Mito Tracker Green FM, and the distribution of mitochondria was observed under laser scanning confocal microscope after immobilization of polyformaldehyde.
Results: 144 of the 284 immature oocytes were cultured in vitro, including 88 GV oocytes and 56.IVM 48h for MI oocytes, 56 GV oocytes reached stage M II, the maturation rate was 63.64%, 50 M I oocytes reached M II stage, and maturity rate of 89.29%.M I oocytes was significantly higher than GV. The difference between the two oocytes was statistically significant (P=0.016).
The distribution of mitochondria in the cytoplasm can be divided into three types: peripheral distribution, semi circumjacent distribution and uniform distribution. In 64.10% (50 / 78) GV oocytes, mitochondria are distributed around the mitochondria. In M I oocytes, mitochondria are evenly distributed in 38.71% (24 / 62), 45.16% (28 / 62) M I oocyte lines The mitochondria were distributed in 75.47% (80 / 106) oocytes in 75.47% (80 / 106) oocytes, and the mitochondria in the oocyte were distributed mainly in the peripheral distribution. The difference was statistically significant compared with that of stage M I and M II oocytes, and the mitochondria in mature oocytes were mostly uniform and unmature. There are also significant differences in oocyte, and it can be seen that there are obvious changes in the mitochondria before and after the maturation of human oocytes, and the distribution of the oocyte from the immature oocyte to the mature oocyte mainly from the peripheral distribution to the mature oocyte.
In the 11 oocytes mature in vivo, there are 8 (72.73%) mitochondria in the mitochondria and 3 peripheral distribution (27.27%). Compared with the mature oocyte in vitro, the distribution of mitochondria in the mature oocyte is the concentration of mitochondria in the central region of the cytoplasm, and the fluorescence intensity is higher than that of the surrounding area.
Conclusions: (1) the distribution of mitochondria in the human oocyte before and after maturation is obviously distributed, and the distribution of the mitochondria from the immature oocyte to the mature oocyte mainly is mainly in the mature oocyte. (2) the distribution of mitochondria in the mature oocytes is slightly different from that in the mature oocytes.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R321.1

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相關(guān)期刊論文 前3條

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本文編號(hào)：2076708