上传用户：icnvnjnjbr文档下载 ：『』&&『』『』学位专业：&关 键 词 ：&&&&&权力声明：若本站收录的文献无意侵犯了您的著作版权，请点击。摘要:（摘要内容经过系统自动伪原创处理以避免复制，下载原文正常，内容请直接查看目录。）人脐血源基质细胞(human umbilical cord blood derived stromal cells，hUCBDSCs)是我科从2001年开端，在多项国度天然迷信基金赞助下率先从人脐带血平分离造就出并已被国际外承认的一种基质细胞。在对造血微情况功效的系列研讨中发明，以hUCBDSCs为滋养层的体外扩增系统对人脐血CD34~+造血干细胞具有显著的扩增感化，体外与hUCBDSCs共造就扩增后的人脐血CD34~+造血干细胞集落构成才能显著加强；对增进巨核细胞集落构成单元(CFU一Mg)的构成具有显著优势。植物试验研讨成果进一步显示，hUCBDSCs经尾静脉输注可敏捷归巢至小鼠骨髓并能历久生计；在对裸鼠造血毁伤修复的研讨中，发明hUCBDSCs具有增进造血毁伤修复的感化。在体外研讨中发明hUCBDSCs对增进巨核细胞集落的构成感化比人骨髓基质细胞加倍显著后，进一步在造血微情况毁伤裸鼠模子中，植入巨核系Hel细胞和人脐血源基质细胞，发明人脐血源基质细胞在体内增进巨核系细胞增殖和修复造血微情况都要强于人骨髓源基质细胞。以上研讨提醒人脐血源基质细胞在增进造血方面较骨髓源基质细胞具有必定稳固性和必定优势，对造血微情况毁伤具有修复感化。绝对于骨髓而言，人脐带血起源普遍，获得便利，不受品德伦理限制。是以，人脐血源基质细胞的发明为临床上放/化疗、造血干细胞移植后或其它血液性疾病改良造血微情况和增进造血恢复带来了新远景。然则，人脐血源基质细胞对增进造血干细胞向红系、粒系、单核系、巨核系、淋巴系分化的详细感化毕竟若何，还没有相干文献报导，须要进一步研讨。今朝，在对基质细胞和间充质干细胞(mesenchymal stem cells， MSCs)的熟悉上，存在很年夜的争议，惹起争议的重要缘由是这两类细胞本身缺少特定的份子标志。依据对人脐血源基质细胞的研讨任务和熟悉，以为基质细胞和间充质干细胞是统一来源，分歧发育阶段的两类细胞。这两类细胞在细胞生物学特征及其造血调控感化有没有差别，有需要停止体系的比拟研讨。据此，本课题拟经由过程比拟人脐血源基质细胞和人脐血源间充质干细胞体外有关生物学特色及其对人脐血CD34~+造血干细胞的粘附率、迁徙率；对人脐血CD34~+造血干细胞引诱分化感化等方面商量来提醒人脐血源基质细胞和间充质干细胞体外支撑、调控造血感化的特色，以丰硕对人脐血源基质细胞和间充质干细胞的周全熟悉，为往后人脐血源基质细胞的临床迷信运用供给实际和试验根据。办法：(一)人脐血源间充质干细胞和人脐血源基质细胞的分别造就及其细胞生物学特征的比拟1。人脐血源间充质干细胞(hUCBDMSCs)和人脐血源基质细胞(hUCBDSCs)的体外分别造就。安康婴儿(足月胎龄)脐带血在4小时内经6%的明胶沉降红细胞后，用1。077g/l的Percoll淋巴细胞分别液，密度梯度离心取得单个核细胞(mononuclear cell，MNC)。所得单个核细胞分离在改进Dexter系统造就hUCBDSCs，在间充质干细胞专门造就基(mesenchymal stem cell medium， MSCM)造就hUCBDMSCs。2。颠倒显微镜下不雅察两类细胞的形状、贴壁、集落构成，细胞舒展及融会情形；采取CCK一8法和Transwell法分离检测两类细胞的体外增殖、粘赞同对hUCB一MNCs迁徙的影响；两类细胞周期及细胞外面抗原的流式检测(细胞外面抗原选择：Fn、Lm、CD45、CD29、CD44、CD34、Stro一1和CD106)。3。两类细胞向成骨、成脂、成软骨细胞引诱分化的检测。用茜素红染色检测成骨情形，用油红染色检测成脂情形，用阿辛蓝染色检测成软骨情形。(二)人脐血源间充质干细胞和人脐血源基质细胞体外调理造血干细胞感化的比拟1。分离以两种细胞为滋养层细胞，对人脐血单个核细胞短时间共造就后，行半固体造就法比拟CFU一GM、CFU一E和CFU一GEMM构成情形，未行共造就的造血干细胞为对比组。2。用逆转录PCR和及时定量荧光PCR办法检测两类细胞中G一CSF、TPO、SCF、SDF一1和GM一CSF细胞因子基因的表达。3。用ELISA法测定造血干细胞分离与两类细胞共造就0、7、14天分歧时相点，上清液中G一CSF、TPO、SCF、SDF一1和GM一CSF细胞因子排泄变更情形。4。将用免疫磁珠分别法所得的脐血CD34~+造血干细胞分离与hUCBDSCs和hUCBDMSCs共造就，用流式细胞仪于共造就第7、14天测定CD34~+造血干细胞分离向粒系、红系、巨核系和淋巴系分化的情形。粒系选用的细胞标志：CD14、CD15、CD33；红系选用的细胞标志：CD71；巨核系选用的细胞标志：CD41a；淋巴系选用的细胞标志：CD3、CD19。CD34~+造血干细胞变更的检测标志仍选用CD34。成果：(一)人脐血源间充质干细胞和人脐血源基质细胞的分别造就及其细胞生物学特征的比拟1。人脐血源间充质干细胞(hUCBDMSCs)和人脐血源基质细胞(hUCBDSCs)的分别造就所取得脐血体积50一100ml，均匀每份84ml，脐血所含细胞数均匀为2。3×10~9/ml，经密度梯度离心能分别出的单个核细胞数均匀为7。0×10~5/ml。每份脐血都可造就出hUCBDSCs，约67%的脐血可造就出HUCBDMSCs。2。 hUCBDSCs和hUCBDMSCs镜下形状不雅察、细胞增殖、迁徙、细胞周期及细胞外面抗原检测脐血源单个核细胞在改进Dexter系统造就hUCBDSCs，3一4天后，粘附在瓶壁的细胞形状有圆形、三角形、纺锤形和不规矩形，造就7天后贴壁细胞重要由巨核样、纤维样和小球样细胞构成。用统一缩小倍数在分歧视野分类和计数细胞，巨核样细胞约占48%，纤维样细胞约占44%，小球样细胞约占8%，细胞融会达80%需30多天。在造就进程中无集落构成，按12的比例传代，传代才能差，最多传到第3代。传代后细胞仍为异形性，跟着传代增长，细胞舒展和融会都显著延迟。传代后前4天细胞处于静息期，4天后开端呈对数性发展，在第8天到达细胞增殖最岑岭，随后增殖才能渐渐降低并进入一个平台期。脐血源单个核细胞在MSCM造就基中造就hUCBDMSCs，在造就前7天，贴壁细胞形状以纺锤形、三角形和小球形多见，但造就10天后，一些纤维样/纺锤体样细胞逐步长成稠密的集落，并开端疾速发展，其它形状细胞逐步消逝，细胞形状趋于统一性，约28天细胞可长满造就瓶，可按13的比例传代，年夜约每5一7天传代1次，可持续疾速扩增细胞12代，细胞形状同原代雷同无变更。年夜约每1×10~8脐血单个核细胞可构成5个纤维样集落。传代后前2天细胞处于静息期，2天后开端呈对数性发展，在第4天到达细胞增殖最岑岭，随后增殖才能降低并进入一个平台期。Transwell法检测两类细胞对等量脐血源MNC的迁徙，hUCBDMSCs组的膜下迁徙细胞数：152±37，hUCBDSCs组的膜下迁徙细胞数为237±22，经配对样本t磨练p=0。03。hUCBDMSCs细胞周期检测，G0/G1期占86。34%，S期占4。14%，G2+M期占9。61%。hUCBDSCs细胞周期检测：G0/G1期占76。18%， S期占9。87%，G2+M期占13。94%。hUCBDSCs和hUCBDMSCs都表达Fn、 CD44和Stro一1，都不表达CD34。hUCBDMSCs表达Lm和CD29，但hUCBDSCs不表达。hUCBDSCs表达CD45和CD106，hUCBDMSCs不表达。3．hUCBDSCs和hUCBDMSCs向成骨、成脂、成软骨细胞引诱分化的检测hUCBDMSCs可胜利引诱向成骨、成脂、成软骨分化，hUCBDSCs不克不及向成骨、成脂、成软骨引诱分化。(二)人脐血源间充质干细胞和人脐血源基质细胞体外调理造血干细胞感化的比拟1。人脐血单个核细胞分离与hUCBDSCs和hUCBDMSCs短时间共造就后，行半固体造就基造就比拟CFU一GM、CFU一E和CFU一GEMM构成情形人脐血单个核细胞分离与hUCBDSCs和hUCBDMSCs短时间共造就5天后，以hUCBDSCs为滋养层共造就的造血干细胞构成CFU一GM数高于以hUCBDMSCs为滋养层组，p《0。01。以hUCBDMSCs为滋养层共造就的造血干细胞构成CFU一GEMM数高于以hUCBDSCs为滋养层组，p《0。05。CFU一E构成数两者无明显性差别。2。 hUCBDSCs和hUCBDMSCs中G一CSF、TPO、SCF、SDF一1和GM一CSF细胞因子基因的表达的PCR检测hUCBDSCs表达G一CSF基因高于hUCBDMSCs，而hUCBDMSCs表达TPO、SCF、SDF一1和GM一CSF基因要高于hUCBDSCs的表达。3。用ELISA法测定造血干细胞分离与两类细胞共造就0、7、14天分歧时相点，上清液中G一CSF、TPO、SCF、SDF一1和GM一CSF细胞因子排泄变更经ELISA检测，上清液中细胞因子G一CSF、TPO、SCF、SDF一1和GM一CSF呈静态的排泄变更。在造就的0天，hUCBDSCs组上清中G一CSF的含量高于hUCBDMSCs组，SCF含量低于hUCBDMSCs组。TPO、GM一CSF和SDF一1的含量两组相当无统计学意义，p》0。05。两组都有细胞因子G一CSF、SCF、SDF一1在共造就第7世界降，在第14天上升的表示。TPO则在两组共造就14天中都表示为连续上升，在0天、7天和14天3个时相点，hUCBDSCs组的排泄量均高于hUCBDMSCs组。GM一CSF在hUCBDMSCs组表示为在第7世界降，在第14天上升；在hUCBDSCs组则表示为连续上升，且在后两个时相点的含量高于hUCBDMSCs组。4。共造就后CD34~+造血干细胞分离向粒系、红系、巨核系和淋巴系定向分化的检测经免疫磁珠分选CD34~+造血干细胞流式检测阳性率为90。2%，共造就7天后两组中CD34~+造血干细胞降低；两组中CD19、CD3和CD41a呈阴性表达，CD33、CD71和CD14呈阳性表达。CD15在hUCBDSCs组呈阳性表达，在hUCBDMSCs组呈阴性表达。CD14的表达hUCBDSCs组高于hUCBDMSCs组；CD33和CD71在两组的表达相当，无统计学意义，p》0。05。在共造就第14天，CD34~+造血干细胞进一步降低；CD19和CD41a在两组的表达仍为阴性，CD15、CD3、CD33、CD71和CD14表达阳性；两组中CD33表达呈上升趋向，CD71呈降低趋向；在hUCBDSCs组CD15的表达虽为上升趋向，但此时排泄量低于hUCBDMSCs组；此时相点hUCBDSCs引诱CD34~+造血干细胞向CD33~+的髓系细胞和CD3~+的淋巴细胞分化要强于hUCBDMSCs；在此时相点，红系表达的CD71在两组都呈显著的降低。结论：1。脐带血经雷同密度梯度离心所得的单个核细胞，用适合的造就基造就可胜利取得hUCBDSCs和hUCBDMSCs，这两类细胞具有分歧的细胞生物学特征，应用hUCBDMSCs具有较多的干细胞特征可以将两类细胞差别。2。 hUCBDSCs在体外晚期增进CD34~+造血干细胞向髓系定向分化要优于hUCBDMSCs。hUCBDSCs在增进造血干细胞向巨核细胞分化和血小板生成方面比hUCBDMSCs具有更年夜的潜能。Abstract:Human umbilical cord blood stromal cells (human umbilical cord blood derived stromal cells hucbdscs) in our department from the beginning in 2001, in a number of National Natural Science Foundation under the auspices of lead from human umbilical cord blood split from created and has been the acknowledged a stromal cells. In the hematopoietic micro environment effect of the series of research invention by hucbdscs trophoblast in vitro amplification system of human umbilical cord blood CD34 ~ + hematopoietic stem cells with significant amplification effect, in vitro and hucbdscs were created after amplification of human umbilical cord blood CD34 ~ + hematopoietic stem cell colony form to sign to promote megakaryocyte colony form unit (CFU mg) has a significant advantage. Plant test research results further show that the hucbdscs by tail vein infusion of agile homing to the bone marrow of mice and long- in the research on nude mice hematopoietic damage repair, the invention of hucbdscs can enhance hematopoietic damage repair action. In vitro study invention hucbdscs to promote megakaryocyte colony of forms action than human bone marrow stromal cells double significantly further in the hematopoietic damage in nude mice model, implantation megakaryocytic HEL cells and human umbilical cord blood stromal cells, inventor of umbilical cord blood stromal cells in vivo enhance megakaryocytic cell proliferation and repair hematopoietic micro are stronger than the human bone marrow stromal cells. Discussion of these remind human umbilical cord blood stromal cells in promoting hematopoietic than bone marrow stromal cells with a certain stability and advantages must, of hematopoietic micro damage has repair effect. Absolute in the bone marrow, the origin of human umbilical cord blood is common, convenient, not restricted by moral and ethical. Is, after the invention of human stromal cells derived from umbilical cord blood for clinical put / chemotherapy and hematopoietic stem cell transplantation or other blood diseases improved hematopoietic micro environment and promote hematopoietic restoration brings a new perspective. However, human umbilical cord blood stromal cells to enhance hematopoietic stem cells into erythroid, granulocytic and monocytic and megakaryocytic, lymphoid differentiation detailed action after all how, there is no coherent reported needs further discussion. Today, in familiar of stromal cells and mesenchymal stem cells (mesenchymal stem cells, MSCs), there is a very big controversy and sparked controversy of the important reasons is the cell itself lacks specific molecular markers. According to the research task of human stromal cells derived from umbilical cord blood and familiar, that stromal cells and mesenchymal stem cells (MSCs) is a unified source and at different growth stages of the two kinds of cells. There is no difference between these two types of cells in the cell biological characteristics and their hematopoietic regulatory effect, there is a need to stop the system of comparative study. Accordingly, this paper intends to through the process than anthropomorphic umbilical cord blood stromal cells and human umbilical cord blood derived mesenchymal stem cells in vitro on biological characteristics and of human umbilical cord blood CD34 ~ + hematopoietic stem cell adhesion rate, on human umbilical cord blood CD34 ~ + hematopoietic stem cells induce differentiation effect and other aspects to discuss to remind human umbilical cord blood stromal cells and mesenchymal stem cells in vitro support, the characteristics of hematopoietic regulation action, with rich of human umbilical cord blood stromal cells and mesenchymal stem cells (MSCs) a comprehensive understanding of, for the descendants of stromal cells derived from umbilical cord blood of clinical science to use actual supply and test according to. Way: (a) from human umbilical cord blood mesenchymal stem cells and human umbilical cord blood stromal cells were created and cell biological characteristics comparison 1. Human umbilical cord blood derived mesenchymal stem cells (hUCBDMSCs) and human umbilical cord blood stromal cells (hUCBDSCs) in vitro were created. Ankang infants (gestational age) umbilical cord blood in 4 hours after 6% of the gelatin sedimentation of red blood cells, with 1. Percoll 077g/l lymphocytes were liquid, density gradient centrifugation to obtain mononuclear cells (cell mononuclear, MNC). The resulting mononuclear cells were isolated in an improved Dexter system to create hUCBDSCs, in which stem cell medium mesenchymal, MSCM, and hUCBDMSCs were created in the mesenchymal stem cells (MSCs). 2. Under the inverted microscopy observations of two classes of cells shape, adherent and colony form, cells spread an take CCK 8 and Transwell separation detection are two types of cell proliferation in vitro, sticky agree with effects on migration of HUCB mononuclear cells (MNCs); two kinds of cell cycle and cell outside the resistance of flow cytometry (outside the cell antigen selection: FN and LM, CD45, CD29, CD44, CD34, stro signing a 1 and CD106). 3. Detection of two kinds of cells to induce differentiation of osteoblasts, osteoblasts and chondrocytes. With alizarin red staining of bone, oil red staining and adipogenic, using alcian blue staining chondrogenic conditions. (two) human umbilical cord blood derived mesenchymal stem cells and human umbilical cord blood stromal cells in vitro conditioning hematopoietic stem cell effect compared with 1. Separation in two kinds of cells cytotrophoblast, on human umbilical cord blood mononuclear cells in a short period of time were created after, semi solid makes the incomparable CFU GM, CFU E and CFU GEMM forming, without co created by hematopoietic stem cells as the contrast group. 2. Detection of G, CSF, TPO, SCF, SDF, 1 and GM a CSF cytokine gene expression in two kinds of cells by reverse transcription PCR and real-time quantitative fluorescence PCR. 3. Determination of hematopoietic stem cell separation and two kinds of fine by ELISA目录:中英文缩写一览表5-7Abstract7-14摘要15-20前言20-23第一部分 人脐血源间充质干细胞和人脐血源基质细胞的分离培养及其细胞生物学特性的比较23-41&&&&材料与方法23-32&&&&结果32-39&&&&讨论39-40&&&&小结40-41第二部分 人脐血源间充质干细胞和人脐血源基质细胞体外调节造血干细胞作用的比较41-70&&&&材料与方法41-51&&&&结果51-66&&&&讨论66-69&&&&小结69-70全文总结70-71致谢71-72参考文献72-80文献综述 人脐血源基质细胞的研究进展80-96&&&&参考文献86-96攻读博士学位期间发表的文章96-97发表论文97-119分享到：相关文献|代孕妈妈与其子女有无血源关系?_百度知道
代孕妈妈与其子女有无血源关系?
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没有血缘关系
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(?) ?盻诀???┦??ぃ??ずだ猚ア秸τま璓逼?钵盽??的?┪?甤ず饯钵?τま璓块?恨超峨┪?的ず翲硈の?ぱ┦?甤挡篶钵盽单?常穦旧璓?┦ぃ▅?????┦????τぃ▅?薄鶪ョ亥镣炊筂?
(⒈) 逼?ア秸??┦?逼?ア秸τ旧璓ぃ▅???κ?ぇ??????ぼ?ぱ?秅戳ㄓ璸衡?璝秅戳?丁┑?┪跑眔ぃ﹚戳?硂陪ボ?衡?逼?ア秸?
(⒉) 块?恨?穕┪超峨??κ?ぇ?????きぇぃ▅?琌??块?恨?穕┪超峨┮璓?筁┕?块?恨も砃┪?的稰琕?甤?ゥ┪?甤ず饯钵?А?旧璓块?恨?穕?τ?的稰琕??⊿Τヴ?紉???诡谋?
(⒊) ?甤ず饯钵??????Τκ?ぇ?き?ぃ▅Θ??琌パ?甤ず饯钵?┮ま璓???甤ず饯钵?痝琌?タ盽?甤ず饯舱麓????甤????よ?
(?)●╧┦???
弘睪ぃタ盽?Τ参璸陪ボ??30%?ぃ▅琌パ╧┦ま癬?ヴ??紇臫弘挛籹硑┪块癳?ぃ▆???
??废?都皊?窤???钵盽?弘舗繰?Ρ眎?ネ崔竟﹛?も砃┪祇?τ?穕单?常??旧璓╧┦ぃ▅????程???╯祇瞷?╧┦ぇ礚弘痝┪弘挛伐祡ぶ?拜肈?讽い场?琌パ?框肚??┮璓?
?????ぃ??
璝冈荷?痜菌の浪琩???????禘琩?А礚猭眔?Τ闽ひ包ぃ▅??絋秆睦?硂贺薄猵参嘿?Θ?ぃ??ぃ▅???摸ひ包玥ㄏтぃ?ぃ▅?????璝?よ???┪ぃ▅?戳禬筁3???ㄤ?礛?ゥ?诀穦ョ穦?碩???
1.4& ぃ▅?溃?
ぃ▅ぃ虫琌ㄓ???ㄓ?溃??癸??癸ひ包ㄓ弧?ネㄠ▅???琌????╧?┪???ōだ??﹚?疭?琌?┦?ゼ?竒菌?る胔璍??钩???┦?┦?疭借?Τ?翴?ろ???硂?籔?┦?ダ┦?闽??镑ㄅ緄???ㄠ????骸ì??ダ?ōだ?讽礛?癸╧┦ㄓ弧?璝ぃ▅?Θ?琌ㄓ?╧┦?ㄒ?弘??计秖ぃì┪┦??ア秸单??穦癸╧┦à︹侥阑????╧┦?Τ?┦?稱┑????ネ㏑㎝蚌▅???ぱ戮?
ひ包ぃ?Τ???ㄠ??ぃ?Θ??ダ?ア??贺⊿Τ???盢ㄓ?谋眔??ぃ镑Ч俱单?Τ?ㄇ膀服畕ひ包??穦??癸?Τ极?????ぃ界褐倒???
⒈?虏ざ瞷さネ崔?м?摸?
2.1& 瞷さ?秈甶
2.1.1& 璍??秘
1994??璣瓣?摸ネ▅籔璅璍?╯恨瞶Ы(HFEA)矗??眖瑈玻?璍??舱麓?ノ??摸???ㄓ方???某???场?チ渤眏疨は癸ぇ??????る??某皘硄筁?窽ゎ璍??秘?
2.1.2 ?Μ緄???璅璍
??砰??弘τ???璅璍?????禞??き?????戳??戳?獽璶砆綪反????璝璅璍??ダ腀種???獽?ㄑ?烩緄?
2.1.3& 礚┦ネ崔
?м砃琌?ノ?だてЧΘ?砰灿璏??琵ウ??Go?篈????筿阑ㄏㄤ籔?????磕??秈τ币笆??穝?璅璍祇▅筁祘?瞷さ??瞶?σ納?既礚瓣產甧砛硂?м徊?ぃ▅ひ包ネ▅?
2.2& ?翠矗ㄑ?徊?ネ崔?м
2.2.1& 护祇逼? (OI) (OI+IUI)
⊿Τ逼??包?硓筁狝ノ┪猔甮縀??玃ㄏ?盻睦?Θ剪????璝?盻??睦??筁?采????獽??穦旧璓Τ?璏璍?薄猵?瞷?
2.2.2& ?甤ず弘睪猔甮
硂よ猭琌??龟喷?ず?眖?ひ矗ユ?弘睪妓セぇい?┾??篈タ盽のㄣ?笆???弘??Θ?蔼緻??弘睪妓セ??皌?ヾ?狝ノ縀?护旧逼???ゐ斗陈綤?薄猵???盢蔼緻??弘睪妓セ猔??甤ず???ㄏノ?よ猭??続??︽┬Τ?螟?╧┦弘睪Τ淮稬拜肈?┪ぃ▅Θ?ぃ??ひ包?
2.2.3& 砰??弘 (IVF)?刚恨捆ㄠ?
?眖?ひのヾ?砰ずだ???弘?㎝???礛??龟喷?ず?р弘?の??挡?Θ?璅璍(?弘?)?ㄤ???盢璅璍???┦??甤ず?琵ウ??甤い?礛Θ??硂よ猭続ノ?块?恨?穕?┪Τ腨??甤ず饯钵??ぃ▅???璝盢硂よ猭挡???璏歼ず虫弘?猔甮??玥???獀励腨??╧┦ぃ▅拜肈???さら?洛厩??礚靡沮陪ボ?刚恨捆ㄠΤ耕?诀穦?瞷?ぱ┦钵盽?薄猵?
2.2.4& 璏借ず猔弘砃 (ICSI)
?龟喷?ず?盢虫?弘??钡猔甮??┾???┦???虑?矗蔼?弘ゑ瞯?硂よ猭?璶?腊?腨???タ盽弘??┪块弘恨?穕?ぃ▅???
2.2.5& 块?恨ず皌?簿从
硂よ猭籔?砰??弘?摸??だ????よ猭琌?も砃?ず?盢??㎝弘???猔?块?恨ず?ㄏ?弘??钡??砰ず?Θ?τ獶?砰??弘??龟喷?ず?Θ?
2.2.6& ㄑ弘???弘
?ノ?甤ず弘睪猔甮?よ猭?盢?弘??弘睪猔??┦?甤ずτ笷?С甒ヘ??硂よ猭続ノ??ひ弘?借?ぃㄎ??笆?耕畓?┪弘?计秖筁????⊿Τ弘?单??
2.2.7& ???秘
盢?ネ▅?包?┮?????籔ぃ?籹硑???包??︸玅?弘睪睼???秈︽砰??弘?
2.2.8& ?瞶胔ゥ
瞷さぇネ崔?м祘???????薄猵?ㄌ沮?ダ?逼τ矗ㄑ?
(?) ノ?赣祘??皌?妮盉?蛮よ???皌??の
(?) 赣琿盉?い?ヾ?ぃ??尿胔ゥ?玻戳??癸?τē??⊿Τㄤ?ち龟?︽?蠢?獀励よ??
⒉?だ猂?摸ネ崔?м徊???瞶?σ
3.1& ???瞶??σ
3.1.1& 璅璍?㏑笲
?沮セ翠??包?洛ネ┮矗ㄑ?戈??硓筁?砰??弘?(IVF)の?璅璍簿从?(ET)?Θ?瞯??κ?ぇ???
τ包???匡拒?м徊?ネ▅?璅璍??计穦钡??贺从玡框肚厩禘耞? (PGD)?縵匡璅璍?旧璓Τ?秖璅璍砆綪反?硂琌?ョ丁钡?躬纘包?糧璍㎡?
????旅?璅璍┪ぃ砆从?τ?緇?璅璍?┕┕砆??躬纘ノ??砰璅璍??╯???硂よ??笵紈?σ?龟?ぃ甧┛跌??
3.1.2& ﹀参??
パ???瞶闽玒??Τ贺贺岿侯狡馒?????猭??ョ螟???﹚?疭??克妮猭の膥┯猭??矪瞶??ぶア?肚参?ㄌ沮?
??ぃ阶琌弘?┪????秘?┪琌??ゥダ??Α把籔徊?ネ▅?м?┘?常Τ帝?硈?闽玒の紇臫?τ┘?丁?闽玒А??盿?ぃ???繧の拜肈??????ゥダ??碞???季▅舦τ癸Τ闽??ダ矗?癬禗??┪砛穦?ゼ眔?癠??ダ?種???︽いゎ胔ゥ??秈︽糧璍︽笆??硂贺贺產畑闽玒??пΡ?碞琌??Τ¨ネ崔ル︸??ネ?〃?瞷┮盿ㄓ??诀?
3.1.3& ?繧
?ゥダ?拜肈???盿ㄓ??螟?稱钩??繧???讽????胔ゥ??ダ?秨﹍玻ネ????ぃ??フ㎝?秆?稰薄?讽ネ?ㄓぇ???ǎゥ▅???る?癌ψ?ê贺骸ì㎝尺??薄狐?Τêㄇネ▅?????烩穦??????ゥダ伐Τ????は??
??厨盡癟???摸ネ崔?м?礟酚?砏ㄒ?ネ???ぃ▅ひ包т?ダ秈︽??徊?ネ崔琌砛?????摸ネ崔?м恨瞶Ы?畊辩醇翬弧?т?ダ???徊?ネ玻玡ゲ斗?灿σ納睲贰?猭ㄒ??胔ゥのネ玻???珹?ダ?程沧局Τ捆ㄠ┮妮舦?安??ダネ玻?ぃ腀盢捆ㄠユ临?〆癠??礚璸?琁?
???盢穦玻ネ??繧琌ㄓ??┦???縀逼?筁祘い??┦ゲ斗砆猔甮?贺嘿?Clomiphene Citrate?媚????縀???盻逼????采????τ????Ωい逼?耕?计秖????旧璓?盻?砰縩??勘等?┮盿ㄓ??繧?珹??瘆?盻ま?祇???盻砾单?????硉逼?ョ?ㄏ包?矗Ν秈???戳?
?ノ浮的描も砃蠢包?矗????筁祘い?ㄏノ?ギ?皐恨????浮场┾?Θ剪????┪?ノ浮的描浪い?も砃?筁祘??旧璓竟﹛稰琕?浮饯?﹀单拜肈?
??皌?(??/弘?)??秘???礚?磷??薄猵??ゲ斗璶┯踞パ??怠戳┮礚猭浪喷?ㄓ???琌?ㄇヘ玡临ゼ??痚痜┮璓?稰琕?繧?
3.1.4& 徊?筁祘?癸?溃?
钡?徊?ネ▅?ひ包ゲ斗┯?伐??み瞶溃????伐ぶ?ひ包??硓筁?Ω?徊??獽??Θ??τ–Ω?祘┮盿ㄓ??辨のア辨?旧璓?徊??ひ包?稰?礘納?Τㄇ??穦眞?ы苳??
3.1.5& ?????舦
??ㄣ称縒ミ???碙腨㎝基???ぃ琌?骸ì?ダ?惠璶τネ??????σ納洛厩徊?ネ▅??и?ぃ虫璶眖?ダ?à??σ納??璶眖璶?ネ????à??σ納????о拒?τ?ダ硂??羘嘿??パ??︽???癸?管???????パ?舦??
??计ひ包浩╣?戴辨?留縡??產????セō皌??秘?ㄆ龟?????祇谋?硂ㄇ戈?礚阶??常穦э跑產???穦癸???芠稰?ョэ跑??癸?ダ?芠稰?┮???????せ???烦玡?琌ぃ穦?笵街琌???痷タ?ダ?硂贺?栋砰?┦篡腇︽???瞶?笵紈?琌?眔?σ??
3.1.6?窥籔?痲
埃??????﹟Τㄤ??瞶?拜肈?眔и???σ???м徊?ネ▅┮疉の?琌胑??竒禣???ぃ琌???诀穦常琌А单?硂硑Θ??狡馒??穦拜肈?
??ゥダ?ぇ腀種??ネ??????ネ㏑緄▅獶﹀絫?ネ㏑?┕┕?璶琌?莉?竒蕾?痲?ヘ?????м徊?ネ▅簍跑Θ??胔ゥ坝穨て??贺贺ぃ笵紈?ㄆ祇ネョ??玻ネ?┪砛徊?ネ▅程沧穦祇甶Θ?Τカ初基??種醚?篈?τ筁?癸痜?矗ㄑ程??眔痲?
3.2&&& 膀服毙ネ㏑?瞶
3.2.1&& ネㄠ▅?斗?ひヾ闽玒い秈︽
?沮竧竒???芠???承??ぃ虫承硑?ぱ?窾???砞﹚??ㄇ?穦??(?厩產嘿ぇ??承?????created order)??沮承?癘材?の材?彻??┮砞ミ????璶?承???琌產畑????–?產畑い?ぃ虫–???ひ┪ヾ?璶腊?ㄤ皌案?Θ?癸よ??ネ︸玅?τ?璶ネ緄渤??筂骸????璓?????钡???獀瞶???ЧΘ?〆癠???恨獀砫ヴ?種孔??產畑埃?Τひヾ??ぇ??﹟璶Τㄠ??τネㄠ▅?硂ン???琌?盉?闽玒い秈︽?籔材??礚闽???ネ▅?琌稲薄ネ??穝祇甶?琌??稲薄?挡垂????ネ▅琌ひヾぇ丁?ㄆ?ぃ莱赣Τ盉?闽玒???材??础?ㄤい?
3.2.2局Τ??琌????界の恨產??癠?
??σ硂?拜肈??и?惠璶?笵??癸ひ包琌???ネ▅???琌???ㄥ?τぃ琌舦?????琌???局Τ??????獶琌??匡拒?┪琌砫ヴ?τ琌????界の恨產??癠??膀服畕粄﹚??ぃ琌妮??ダ??τ琌妮?????珿???局Τ舦?ぃ虫?ネ???┪琌﹀絫?à?ㄓ﹚管??タ????琌????界?????ぃ??и??匡拒?┪琌и??北?ぇ??
и?璶絋粄胔ゥネ?琌??┦局Τ?膀セ舦??眖承1:28い笵??虑盉?界倒?摸?甝??硂ぃ琌癸–?癸ひ包?莱砛?τ琌癸俱砰?摸?莱砛??????м徊?ネ▅ぃ虫琌??洛厩拜肈?ョ疉の???舦拜肈???琌瑿㎝地┮界?玻穨?眖??à??????ダ???Θ琌???㏑?狝ㄆ????硂?㏑ぃ?﹚羬?–????
3.2.3&竧竒い璍ㄠ???
??贺从玡框肚厩禘耞(PGD)縵匡?ぃ▆璅璍?旧璓Τ?秖璅璍砆綪反?硂盿ㄓ?拜肈碞琌??┏ネ㏑眖??秨﹍?璝眖钢22:9-10?139:11-16?瑿1:5??1:15?隔1:15?璅璍???Τネ㏑琌?∕???龟?ぃ琌?洛厩à?﹚??
ゼ?ネ?璍ㄠ琌虑????硑Θ???廷┮?ǎ??跌??皌砆??㊣酬??箇戳???ネΘ????瓣?㎝????琌ゼ?ネ??硑???廷??倒??????(??3-4?钢??き6-7???3????73?辽???2?24)?┮?﹟??甤い???﹟ゼΘ????ㄣΤ基??硂琌?癸璍ㄠ??猭?
3.2.4ダ克籔浮い???闽玒
胔ㄈ疭(John
Wyatt)??狦и?粄痷??承硑???ê或???Τ绊眏?瞶パ??蝴臔Θゥ㎝ゥ▅?闽玒?浮い???蛤ダ克ぇ丁?ê?稲钡??闽玒??﹚琌承硑????场???酚??承硑砞璸?ダ克р??╆?胔い????钡??ぃ琌?ネ??τ琌?竒抄????る?ネ㏑?が獺が稲?闽玒?竒?ミ??┮??瞶胔ゥ硂贺м砃??セ?э跑???砞璸?
3.2.5& 碙腨?基?
&&&&& 坝?Α?ネ▅筁祘??穦??搭???硑??ê?????碙腨??imago????碙腨琌??┮结ぉ???┮Τ?А?ㄉΤ??τぃ穦??穦???τΤ┮ぃ???膀服畕ひ包??碝―?產畑い⊿Τ???ぃЧ骸?ぃ莱虫眖洛厩?????碝―腊????莱赣????籔?Τ?盞ち?闽玒τ??????硂妓?琌碝―??Ч骸?ネ程?璶?よ??
3.2.6 ぱ?毙?瞶阶?
?贝癚Τ闽ネ崔徊???瞶拜肈??ぱ?毙??猭矗ㄑ?獶盽?璶?阶翴??翠そ毙盉?徊旧穦羆稦ㄆ?ゅ眃眖ぱ?毙獺ヵà??莱?玥??弧??ネ㏑ㄓ?ひ包稲?︽?????ゥм砃琌рネ㏑??Θ?玻珇??管?┦?碙腨??
???瓣地┎ЧΘ﹙畒盉?籔產畑厩皘盉??厩揭祘??ゅ眃?るせら钡?セ厨砐拜?眏秸?矪瞶ネ崔?м?拜肈??癸ネ㏑?篈????璶??ま瓃?ぱ?毙毙瞶?嘿?ぱ?琌ネ㏑??甠???琌ぱ???界τ獶莱????弧??ひ包?┦︽???癸よず?瞏ち?????笷倒ぉ㎝钡??ō砰粂ē????ゥ獽琌笻は??瞶種竡??
?ゅ眃尿弧?毙穦?獶?絃は癸ネ崔?м?ㄤ龟??м籔獺ヵ??狝叭?竤??種竡?⊿Τ侥???????ネ崔?м????Τネ▅?螟?ひ包????礛產畑璸购??包?洛ネゴ硄块?恨┪块弘恨??の狝?琘ㄇ媚?单?礛よ猭??瓃よ猭琂?糤?ヾ?胔ゥ?诀穦?ョぃ笻は毙穦碙??摸ネ㏑㎝碙腨??玥?
??蹦ノ┦?匡拒?磷?框肚痜??ゅ眃?ボ?ひ包莱獺ヴぱ???逼?ぃ???块ゴ墓璶????み篈?北?ネ㏑??玥獽籔毙穦は癸?纔ネ厩礚钵???ぃ▅?ひ包??ん?ボ赣穦穦徊旧??钡?ぱ?┮界???琜??辨璶?????σ納烩緄狝叭?
⒊??м????隔
4.1& ?界ネ㏑??毛―
????莱す?蓟?毛??┮??κ?碞胔ゥ??承??き21??ㄤ龟??绊?ぃ居?硂妓毛??竒?????癸酚承??き20㎝??き26??硂??獵??ひ┯脓???克ㄈ?┰╱?獺み?瑻み?单???界???璢盖?
4.2& Μ緄ㄠ?
?緄??琌ㄏ??????眔??矪?ê琌??玡?砆??管??タ???ぃ▅?馣び┮ē??籔ㄤぃ耞钡?徊?筁祘?礹璚籔溃????硑Θぃぶ璅璍ネ㏑?犠???ぃ緄▅硂ㄇ单稲????膀服畕ひ包Μ緄???琌р⊿Τ產???盿秈??產????孔??緄▅籔??框肚礚闽?????瞏砰穦?孔礚╬?稲??┮???籔?ダ?﹀方闽玒??ぃ琌程?璶?Μ緄??讽礛琌??Τ獀励┦の??????癸ぃ▅?よ猭?τΜ緄??癸?膀服畕??????紇臫???癸?穦??弧琌κ?τ礚?甡?
场??盉?闽玒?瞷拜肈┪?诀??钩ㄢ竲慈??ぃ?竲?惠璶材?唉竲?や嫉のキ颗?讽礛????絋癸?ダ闽玒癬妒てのタ???ノ??安璝ㄢひ包?闽玒Τ帝?方?拜肈?碞讽眖?セ秆∕???糧?產畑獀励?畍ㄓ宾?腳冻?Murray BowenFamily Triangulation
??盉??種竡の祇甶よ??????????穝?逼?贺ネ?ㄆン?纔?┦?秸俱┪璹ミ????產畑?ㄏ㏑?
5.1& ??醇紌?о拒
и?惠璶?フ??мネ▅?⊿Τ痷タ秆∕ぃ▅?ぃ▅?包??琌虑?м腊???ノ獶?礛?よ猭Θゥ?传?杠弧?ぃ▅?包?ご礛ぃ?镑钩??ネ▅包??繦種Θゥ????癸?癸кぃ▅?и???ノ?┮Τぃ??よ猭??籔????и??沽刚?ノ?м?搭?みい??踞ぇ??и?璶?辆?のг瑻?み?腨タ??癸?贺?闽玒??み?颗秖?贺?匡拒??ノ毛??み???贺?∕﹚??ゼ蹦?ヴ?徊?ネ▅よ猭???и?讽礛惠璶瞏??╯?贺?︽┦?よ猭??フ讽い?笵紈瞶???繧?????????ゲ斗?碝―???み種?
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