{"created":"2023-06-19T07:18:09.644277+00:00","id":3281,"links":{},"metadata":{"_buckets":{"deposit":"b2e92eb3-4530-4f59-9041-81538595d938"},"_deposit":{"created_by":4,"id":"3281","owners":[4],"pid":{"revision_id":0,"type":"depid","value":"3281"},"status":"published"},"_oai":{"id":"oai:az.repo.nii.ac.jp:00003281","sets":["370:15:392"]},"author_link":["16384","16383"],"item_10006_date_granted_11":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"1985-10-02"}]},"item_10006_degree_grantor_9":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_name":"麻布大学"}]}]},"item_10006_degree_name_8":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"獣医学博士"}]},"item_10006_description_22":{"attribute_name":"Abstract","attribute_value_mlt":[{"subitem_description":"Abstract\n Cell culture initiated in 1907 by Harrison's nerve fiber culture has achieved so tremendous a progress as to enable detailed in vitro studies using cultured cells.\n In this study, culture of granulosa cells (GC) and luteal cells (LC) from a cow ovary was undertaken to investigate the in vitro behavior of these cells free from the biological control mechanism which is presumed to regulate their behavior in vivo to a certain extent under the control of cerebroneural system, and also their behavior in the presence of gonadotropin (GTH) and prostaglandin F_2α (PGF_2α). The behavioral aspects mainly studied were the multiplication and morphological characteristics of these cells together with secretion of progesterone and estrogen into the culture fluid.\n\nMaterials and Methods\n GC was collected from the follicles of slaughtered animals and from those of live animals of the estrous stage using an injector. LC of the almost functional luteal stage was collected from slaughtered animals. For both the cells, subculture and primary culture cells were used in the experiment. Used in addition were slices of the corpus luteum.\n The culture medium used was an Eagle MEM (Nissui) (1). After dissolution of this medium to a desired concentration, 10% cow and newborn calf sera were added. Culture was carried out by stationary incubation at 37℃.\n GTH consisted of human chorionic gonadotropin (HCG), pregnant mare serum gonadotropin (PMS), prolactin, follicle stimulating hormone (FSH) and luteinizing hormone (LH).They were added to the medium at the commencement of culture. The cell multiplication was determined when the cell growth was microscopically found to be spread over the entire bottom of the culture vessel. The supernatant was freeze-preserved for quantification of hormones. The cells adhered to the culture vessel were counted after the required treatment.\n The cells grown on the bottom of the culture vessel was immediately examined using an inverted phase-contrast microscope. Furthermore, the cells adhered on to the cover glass placed in the culture vessel were, when necessary, stained by Giemsa or May-Gruenwald Giema to examine the cell morphology.\n\nResults\n1. Cell culture of GC and LC\nMultiplication and morphology: GC and LC achieved smooth multiplication in vitro. The cells grown showed an epitheliumlike morphology and indicated a possibility of subculture. Hormone production: Hormone production in the culture fluid seemed to be most affected by the ovarian cycle at the time of collection.\n In culture of GC from animals of the estrous stage, the maximum productins of estradiol (E_2) and progesterone were considerably high, indicating significant differences compared to culture of GC from slaughtered animals. High production of progesterone suggested a functional shift of GC into LC.\n In culture of LC, estrogen production was barely detected. Although progesterone production was large, remarkable variations such as seen in GC were not found.\n2. Response of GTH to GC and LC\nSingle mixture of hormones: HCG, PMS, prolactin, FSH or LH generally accelerated multiplication of GC and LC. Especially prolactin demonstrated a higher accelerative effect than HCG. However, PMS had obviously an inhibitory action on multiplication of GC and LC. \n No demonstrable results were obtained in relation to hormone production.\nCombined mixture of hormones: Within the concentrations used in this experiment, no synergic effect was demonstrated on multiplication of GC and LC. No distinct results were also obtained on hormone production.\n3. Response of PGF_2α to GC and LC\nCell multiplication: PGF_2α seemed to somewhat inhibit multiplication of GC and stimulate that of LC.\nHormone production: Progesterone production was markedly accelerated in GC from slaughtered animals. However, no specific tendency in hormone production could be defined in GC from the follicles of animals at their estrous stage. Estrogen production seemed to be slightly accelerated in 5 of 8 culture vessels of GC from animals of the estrous stage.\n Progesterone production tended to increase in LC. Estrogen production showed no distinct tendency. Almost the identical results were obtained in the experiment of the sliced corpus luteum.\n The above findings confirmed that the phenomena occurring in vitro do not necessarily correspond with biological reactions occurring in vivo and suggested that the sexual function is governed in vivo by a complexity of factors. Thus the experiment with the cultured cells of follicular granulose cells and luteal cells seemed to be a helpful technique that allows simple analysis of the reproductive phenomena occurring in vivo under complicated control mechanisms."}]},"item_10006_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"1907年Harrisonの神経線維の培養に始まる細胞培養は，近年，著しく進歩し，in vitroで培養細胞を用いて詳細な研究ができるようになった。\n　本論文の目的は，乳牛の卵巣からの顆粒層細胞(GC)及び黄体由来細胞(LC)の細胞培養を試み，生体内では上位からの支配を受けて，ある程度その動きが規制されていると考えられているこれら両細胞を生体外にとりだし，フリーの状態にした場合，さらにGC，LCの培養液中に性腺刺激ホルモン(GTH)やプロスタグランジンF_2α(PGF_2α)を添加した場合にどのような態度を示すかを知ることであり，主な観察点としては，細胞の増殖状況や形態などの所見，併せて培養液中へのプロジェステロン，エストロジェン分泌状況の両者について検討を行ったものである。\n　実験材料ならびに方法;GCは，屠場材料及び発情期の生体の卵胞から注射器で直接採取し，LCは，屠場材料でほぼ開花期に近い大きさの黄体由来で，これら両細胞の継代細胞，初代細胞を供試したが，一方，これとは別に黄体をスライスしたものも供試した。\n　培養液は，イーグルMEM培地「ニッスイ」(1)を所定の濃度に溶解後，成雌牛及び新生仔牛血清を10%の割合に添加したもので，37℃の孵卵器で静置培養した。\n　供試GTHは，人絨毛性性腺刺激ホルモン(HCG)，妊馬血清性性腺刺激ホルモン(PMS)，プロラクチン(Prolactin)，卵胞刺激ホルモン(FSH)，黄体形成ホルモン(LH)等で，培養開始時に添加し，細胞の増殖状態を算定する時期は，同じ実験群の中でどれかが鏡検して，ほぼ培養瓶の底面全般に増殖したと見られる時期を選び，上澄はホルモン定量用として凍結保存し，一方，培養瓶に付着した細胞に対しては，一定操作を施して細胞数を算定した。\n　細胞の観察は，倒立位相差生物顕微鏡で，培養瓶の底面に発育増殖した生材料を速やかに観察するとともに，培養瓶内にあらかじめ入れておいたカバーグラスを適時とりだし，これに付着している細胞をギムザ及びメイ・グリュンワルドギムザ染色等を施して細胞の形態を観察した。\n　本研究の結果は，以下のとおりである。\n　1.GC，LCの細胞培養\n　増殖，形態等;GC，LCは，in vitroでよく増殖し，増殖した細胞は，上皮様形態を示し，継代培養も可能である。\n　ホルモン産生;培養液中へのホルモン産生は，採取時の卵巣周期に最も影響されると思われる成績が得られた。\n　GCでは，発情期由来のものは，ちなみにEstradiol(E_2)ならびにプロジェステロン産生量の最高値は極めて高く，屠場材料のものと大差がみられた。なお，プロジェステロン産生量の高かったものは，GCの黄体細胞への機能的推移が示唆された。\n　LCでは，エストロジェンは辛うじて検出できる程度であり，プロジェステロン産生量は高かったが，GCにおけるような大きい変動はみられなかった。\n　2.GTHのGC，LCに対する反応\n　単独添加の場合;HCG，PMS，Prolactin，FSH，LHの各添加は，一般にGC，LCの発育増殖を促進し，とくにProlactinはHCG以上の作用がみられたが，PMSはGC，LCの増殖に対しては，明らかに抑制的に作用した。\n　ホルモン産生については，明瞭な結果は得られなかった。\n　混合添加の場合;増殖については，GC，LCとも本研究での添加範囲内では，GTH相互の協力作用は認められなかった。また，ホルモン産生についても明瞭な結果は得られなかった。\n　3.PGF_2αのGC，LCに対する反応\n　増殖;GCに対しては，増殖を多少抑制し，LCに対しては若干刺激するように思われた。\n　ホルモン産生;GCでは，屠場材料のものに対しては，プロジェステロン産生を顕著に促進したが，発情期卵胞由来のものは，8例全例中5例に産生をやや促進したと思われる結果が得られた。\n　LCに対しては，プロジェステロンを促進するような傾向がみられた。エストロジェン産生については，明瞭な傾向は把握できなかった。黄体のスライスを用いた場合もほぼ同じ結果が観察された。\n　以上の結果，in vitroにおける所見が必ずしもin vivoで観察される臨床的反応とは一致しない現象も確認され，in vivoでの複数の因子による性機能の運営等が示唆される結果も得られる等，卵胞及び黄体由来のGC，LCの培養細胞を用いての実験は，複雑な支配機構によって営まれている生体での繁殖関連現象を単純化して，解析的にこれらを究明する上に有効に利用できる手法と考えられる。"}]},"item_10006_dissertation_number_12":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"乙第232号"}]},"item_10006_version_type_18":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_ab4af688f83e57aa","subitem_version_type":"AM"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"川上, 静夫"}],"nameIdentifiers":[{}]},{"creatorNames":[{"creatorName":"Kawakami, Shizuo","creatorNameLang":"en"}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2013-02-21"}],"displaytype":"detail","filename":"diss_dv_otsu0232.pdf","filesize":[{"value":"61.2 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とくに性腺刺激ホルモン，プロスタグランジンF_2α等に対する顆粒層細胞及び黄体由来細胞の反応について","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"「細胞培養法による牛卵巣機能の検討」 : とくに性腺刺激ホルモン，プロスタグランジンF_2α等に対する顆粒層細胞及び黄体由来細胞の反応について"},{"subitem_title":"A study on bovine ovarian function by cell culture : with special reference to response of granulosa cells and luteal cells to gonadotropin and prostaglandin F_2α","subitem_title_language":"en"}]},"item_type_id":"10006","owner":"4","path":["392"],"pubdate":{"attribute_name":"公開日","attribute_value":"2013-02-21"},"publish_date":"2013-02-21","publish_status":"0","recid":"3281","relation_version_is_last":true,"title":["「細胞培養法による牛卵巣機能の検討」 : とくに性腺刺激ホルモン，プロスタグランジンF_2α等に対する顆粒層細胞及び黄体由来細胞の反応について"],"weko_creator_id":"4","weko_shared_id":4},"updated":"2023-06-19T08:19:47.879944+00:00"}