干细胞在子宫内膜损伤后修复的研究进展

子宫内膜修复是妇产科常见的病理生理过程,分为子宫内膜生理性修复及损伤后修复。生理性修复是内膜自我剥脱与再生的过程,并不形成瘢痕,而损伤后修复常不能完全修复,当损伤因子伤及子宫内膜基底层或破坏子宫内膜生长的微环境时,就会造成子宫内膜增殖受损、内膜再生障碍,甚至形成瘢痕,发生宫腔粘连,影响受精卵或胚胎的植入及着床,导致女性不孕或反复流产。目前,如何促进子宫内膜损伤后修复仍面临巨大挑战。近年来,干细胞对子宫内膜修复的研究备受关注,现就子宫内膜损伤的病理生理及干细胞在子宫内膜损伤后修复的研究进展做简要综述。     

干细胞治疗女性压力性尿失禁的临床进展

压力性尿失禁在各年龄段妇女普遍存在,严重影响女性生活质量及社会形象,现行的治疗手段效果不理想.对压力性尿失禁发病机制的探索,已认识到中段尿道及尿道外括约肌是尿控关键.近年,自体干细胞移植作为新的治疗手段,对女性压力性尿失禁的治疗获得可喜成果.综述干细胞治疗女性压力性尿失禁的近况.     

脐血干细胞的生物学特性及其应用

脐血于细胞为干细胞移植提供了新的选择,与外周血及骨髓干细胞相比,脐血干细胞移植后的低急慢性移植物抗宿主病(GVHD)发病率、低移植相关死亡率(TRM)及疾病复发率具有明显优势,易获得性及免疫耐受等特性使其越来越受到人们的关注,并吸引人们不断进行深入的研究.归纳总结脐血干细胞与胚胎干细胞、骨髓干细胞及外周血干细胞的优缺点...     

干细胞与子宫内膜异位症发病机制的研究进展

子宫内膜异位症（endometriosis,EMs）在育龄妇女中的发病率近15%,其发病机制一直是研究的热点,经典的“经血逆流与种植学说”不断受到挑战,无法解释如盆腔外远处病灶及特殊部位异位病灶的发生等问题,而干细胞学说尝试从“一元论”角度解释所有类型异位病灶的发生。参与EMs异位病灶的干细胞可能来源于子宫内膜、骨髓、卵巢及其他来源。而雌激素、趋化因子、表皮生长因子、血小板源性生长因子等多种因子参与成体干细胞的分化。微小RNA水平与表观遗传学改变可能参与调控间充质干细胞向子宫内膜样细胞的分化过程。异常定植的干细胞可能改变子宫的容受性,导致不孕、流产等不良生育结局。在形成的不同类型EMs病灶中,腹膜型与深部浸润型病灶干细胞基因表达相似,而与在位内膜干细胞基因表达存在差异。随着研究的不断深入,干细胞在妇产科领域的应用越来越广,在治疗卵巢功能早衰、Asherman综合征、EMs相关性不孕及靶向药物研发方面前景广阔。     

干细胞及其外泌体治疗早发性卵巢功能不全的研究进展

早发性卵巢功能不全(premature ovarian insufficiency,POI)是导致女性不孕的重要原因,严重影响患者的身心健康。POI发病原因复杂,近年来发病率逐年上升,且发病年龄趋于年轻化。临床主要采用激素替代治疗,尚无有效根治方法。目前,多种类型的干细胞及其外泌体(exosomes)已应用于治疗POI临床前的实验研究中,并取得一定疗效。干细胞可分为胚胎干细胞和成体干细胞,胚胎干细胞因其伦理争议在临床治疗中受到限制。外泌体可由干细胞分泌,并可携带微小RNA(miRNA)、长链非编码RNA(lncRNA)、生长因子等多种物质,发挥与亲代细胞相似的生物学功能。总结干细胞及其外泌体在POI治疗方面的进展及其可能存在的作用机制,提出其在该领域新的发展前景。     

妊娠晚期羊水间充质干细胞生物学特性研究

目的：研究妊娠晚期羊水来源的胎儿间充质干细胞（MSCs）的体外分离、培养及其生物学特性。方法：贴壁培养法培养妊娠晚期羊水中的胎儿MSCs，形成细胞集落后用细胞刮刀刮下细胞重新培养、扩增，用倒置相差显微镜观察细胞形态．通过细胞表面抗原、细胞核型分析、细胞因子检测等证实该细胞是否为胎儿MSCs，以及不同代细胞之间是否有生物学特性上的差异，并通过绘制生长曲线，比较各代细胞的生长趋势。结果：用贴壁培养法分离、培养出的细胞呈梭形，不同代胎儿MSCs表面抗原、细胞核型相同，不同羊水标本来源的胎儿MSCs均表达胚胎干细胞相关蛋白（Oct-d），第8代（P8代）细胞比P2、P5代细胞生长缓慢。结论：通过上述方法可以成功分离、培养妊娠晚期羊水来源的胎儿MSCs，不同代细胞间生物学特性基本相同。     

干细胞治疗卵巢早衰的研究进展

卵巢早衰（POF）是由多种病因导致的卵泡功能衰竭。大约80%患者属于特发性POF。POF治疗棘手,近年国内外取得了应用干细胞恢复卵巢功能及生育力等方面的动物试验成果,为POF患者卵巢功能及生育功能的恢复带来了希望。干细胞包括胚胎干细胞和成体干细胞,胚胎干细胞治疗因其引起伦理争议而受到限制。目前成体干细胞治疗包括生殖干细胞、骨髓间充质干细胞和胎儿间充质干细胞的治疗。干细胞可能通过分化为卵母细胞或通过旁分泌抑制卵泡凋亡来修复受损卵巢。现对POF的干细胞治疗进展进行综述。     

Stem cell research for male infertility

Stem cells have the ability both to differentiate into numerous tissues and to self-renew. Because of these unique properties, stem cells are promising candidates for use in regenerative medicine. Among stem cell types, embryonic stem (ES) cells have been the most studied; however, alternatives such as induced pluripotent stem cells or other adult stem cells are now being established. In this review, we focus on stem cell research that may have applications in treating male infertility. Stem cells with ES-like properties have been generated from adult human testis tissue. We expect that breakthroughs in stem cell research will increase our understanding of male infertility and lead to treatments in the near future.     

OCT4在妇产科中的研究进展

八聚体结合因子4(OCT4)是秀丽隐杆线虫神经(POU)转录因子家族的一员,主要表达于胚胎干细胞、生殖细胞、胚胎性癌细胞和肿瘤干细胞中,被公认为是干细胞的标记物,在维持干细胞的多能性、自我更新能力及在肿瘤的恶性行为学中起着举足轻重的作用。同时OCT4也参与胚胎发育过程中多向性分化调节。近年来随着干细胞理论及其相关研究的发展,OCT4作为干细胞的标记物也被用于不同领域的研究中,综述其在妇产科领域的研究进展。     

干细胞在宫腔粘连治疗中的应用

子宫内膜是子宫的重要组成部分,是胚胎着床的场所。子宫内膜可因感染、流产、过度刮宫等因素造成子宫内膜基底层损伤引发宫腔粘连,导致患者月经异常、复发性流产或不孕,严重影响女性的生活质量和生育。然而,目前对于宫腔粘连的各种治疗方法,疗效一般、复发率高。干细胞在再生医学中的广泛应用,使得干细胞疗法成为宫腔粘连的潜在治疗方法,即干细胞可用于子宫内膜的修复和再生。各项临床前实验已经证明,干细胞可减少动物模型受损子宫内膜纤维化,增加腺体数量,提高子宫内膜厚度。综述干细胞治疗宫腔粘连的理论基础、干细胞及其衍生物治疗宫腔粘连的研究进展。     

支持细胞对体外培养精原干细胞的作用途径研究

目的:探讨支持细胞对体外培养精原干细胞的作用途径。方法:选用7 d龄雄性昆明种小鼠,两步酶消化法获得睾丸组织细胞悬液,差异时间贴壁法分离精原干细胞和支持细胞,免疫荧光法和油红O染色法分别对其进行生物学鉴定,流式细胞仪对精原干细胞进行纯度分析。按培养条件的不同将实验分为3组:精原干细胞与支持细胞共培养组(A组)、条件培养基组(B组)、常规培养基组(C组)。其中条件培养基按单纯支持细胞培养清液∶双倍浓缩的DMEM/F12∶胎牛血清=4.5∶4.5∶1的比列配置;常规培养基即含体积分数10%胎牛血清的DMEM/F12。台盼蓝法测定各组贴壁率,四甲基偶氮唑盐(MTT)法测定各组精原干细胞的吸光度并绘制增殖曲线。倒置显微镜下观察各组精原干细胞增殖特点及集落形成情况。比较各组精原干细胞24 h贴壁率、增殖曲线及存活时间的不同。结果:A组精原干细胞24 h贴壁率大于B组及C组(P<0.05),而B、C组间无差异(P>0.05);A组精原干细胞接种起即稳定增殖,于7~10 d形成稳定集落并维持约30 d的特性。B组和C组均表现为精原干细胞经短暂的增殖后呈现快速减少的趋势,培养1周后,精原干细胞数目明显减少。结论:支持细胞对体外精原干细胞的作用是依靠两者之间的直接联系和支持细胞的旁分泌2种途径;而仅依靠支持细胞的旁分泌作用不能促进精原干细胞的贴壁和增殖。     

Eligibility of Cryopreserved Human Embryos for Stem Cell Research in Canada

Objective: To determine whether cryopreserved embryos in Canadian in vitro fertilization (IVF) clinics available (in theory) for research use are actually eligible (in practice) for stem cell research in accordance with the Assisted Human Reproduction Act, the Canadian Institutes of Health Research’s Human Pluripotent Stem Cell Research: Guidelines for CIHR-Funded Research, and the Tri-Council Policy Statement: Ethical Conduct for Research Involving Humans.Methods: Copies of current and past patient information sheets and consent forms for embryo cryopreservation, the future disposition of cryopreserved embryos, and the donation of cryopreserved embryos for research were requested from Canadian IVF clinics. Fourteen of 24 clinics (response rate 58%) provided one or more of the requested documents.Results: A review of the documents currently in use showed that, as of May 2005, all 14 responding clinics require an advance directive for the eventual disposition of cryopreserved embryos. Ten of these 14 clinics allow the donation of cryopreserved embryos for research. Only three of these 10 clinics, however, satisfy in writing the disclosure requirements for embryo research at the time of initial consent for the cryopreservation and future disposition of embryos. Only one clinic specifically identifies the option of embryonic stem cell research.Conclusion: Only three of the responding Canadian IVF clinics provide written details about future donation of cryopreserved embryos for research use and about the need for re-contact in accordance with relevant legislation and guidelines. Two of these three clinics, however, include disposition options that may not be legally available. In the event of death or loss of contact, it will not be possible to get project-specific research consent from the legitimate embryo providers as required by law. These same two clinics also fail to mention embryonic stem cell research as one of the research categories. It follows that we have identified only one Canadian IVF clinic with cryopreserved embryos unequivocally eligible for stem cell research, provided the forms we reviewed were used to obtain consent for the cryopreservation of those embryos.     

Gynecologic Care for Pediatric and Adolescent Patients Undergoing Hematopoietic Stem Cell Transplantation

Hematopoietic stem cell transplantation is used to treat many chronic and acute malignant and nonmalignant conditions. We review hematopoietic stem cell transplantation and its effect on the gynecologic health of pediatric and adolescent patients, including pretransplantation evaluation, contraception, menstrual suppression, sexual health, fertility, primary ovarian insufficiency, and graft vs host disease. Comprehensive and team-based care provides optimal anticipatory counseling, evaluation, and management of acute and ongoing gynecologic issues.     

Development of a xeno-free non-contact co- culture system for derivation and maintenance of embryonic stem cells using a novel human endometrial cell line

Purpose

Mouse embryonic fibroblast feeder layers (MEF) have conventionally been used to culture and maintain the pluripotency of embryonic stem cells (ESC). This study explores the potential of using a novel human endometrial cell line to develop a non-xeno, non-contact co-culture system for ESC propagation and derivation. Such xeno-free systems may prove essential for the establishment of clinical grade human ESC lines suitable for therapeutic application.

Methods

A novel line of human endometrial cells were seeded in a 6-well dish. Filter inserts containing mouse ESCs were placed on these wells and passaged 2–3 times per week. Inner cell masses derived from mouse blastocysts were also cultured on transwells in the presence of the feeder layer. In both cases, staining for SSEA-1, SOX-2, OCT-4 and alkaline phosphatase were used to monitor the retention of stem cells.

Results

ESC colonies retained their stem cell morphology and attributes for over 120 days in culture and 44 passages to date. Inner cell mass derived ESC cultures were maintained in a pluripotent state for 45 days, through 6 passages with retention of all stem cell characteristics. The stem cell colonies expressed stem cell specific markers SSEA-1, Sox 2, Oct-4 and alkaline phosphatase. Upon removal of the human feeder layer, there was a distinct change in cell morphology within the colonies and evidence of ESC differentiation.

Conclusions

Human feeder layers offer a simple path away from the use of MEF feeder cells or MEF conditioned medium for ESC culture. Furthermore, indirect co-culture using porous membranes to separate the two cell types can prevent contamination of stem cell preparations with feeder cells during passaging.