Name: human blastocyst biopsy and vitrification
Uploaded: 2019-07-26T12:30:00
Description: Watch this Scientific Journal Video about Human Blastocyst Biopsy and Vitrification at JoVE.com

AG 和 RM 收集了数据并起草了手稿。DC 分析数据,起草代表性结果,进行统计并修订稿件。FMU 和 LR 对结果和整个手稿进行了批判性讨论。

这里描述的人类胚泡活检方案遵循了G.EN.E.R.A.人类研究伦理委员会的指导方针。
注:有关所需材料,请参阅材料表。所需材料包括实验室鞋类和服装、外科面罩、头发罩、手术手套、永久无毒标记物、钳子和消毒剂。使用手术服,一次性手术手套,面罩,头发盖是强制性的,以防止污染的风险。在开始任何程序之前,必须使用实验室消毒剂(例如 Oosafe)彻底清洁?...

<ol>
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K., Schoolcraft, B., Jansen, R., Mortimer, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Towards Reproductive Certainty: Infertility and Genetics Beyond 1999. , 377-388 (1999).</li><li>Cimadomo, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inconclusive+chromosomal+assessment+after+blastocyst+biopsy:+prevalence,+causative+factors+and+outcomes+after+re-biopsy+and+re-vitrification.+A+multicenter+experience.">Inconclusive chromosomal assessment after blastocyst biopsy: prevalence, causative factors and outcomes after re-biopsy and re-vitrification. A multicenter experience.</a> Human Reproduction. , (2018).</li><li>de Boer, K. A., Catt, J. W., Jansen, R. P., Leigh, D., McArthur, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Moving+to+blastocyst+biopsy+for+preimplantation+genetic+diagnosis+and+single+embryo+transfer+at+Sydney+IVF.">Moving to blastocyst biopsy for preimplantation genetic diagnosis and single embryo transfer at Sydney IVF.</a> Fertility and Sterility. 82 (2), 295-298 (2004).</li><li>Capalbo, A., Rienzi, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mosaicism+between+trophectoderm+and+inner+cell+mass.">Mosaicism between trophectoderm and inner cell mass.</a> Fertility and Sterility. 107 (5), 1098-1106 (2017).</li><li>McCoy, R. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+Selection+against+Complex+Mitotic-Origin+Aneuploidy+during+Preimplantation+Development.">Evidence of Selection against Complex Mitotic-Origin Aneuploidy during Preimplantation Development.</a> PLoS Genetics. 11 (10), e1005601 (2015).</li><li>Scott, R. T., Upham, K. M., Forman, E. J., Zhao, T., Treff, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cleavage-stage+biopsy+significantly+impairs+human+embryonic+implantation+potential+while+blastocyst+biopsy+does+not:+a+randomized+and+paired+clinical+trial.">Cleavage-stage biopsy significantly impairs human embryonic implantation potential while blastocyst biopsy does not: a randomized and paired clinical trial.</a> Fertility and Sterility. 100 (3), 624-630 (2013).</li><li>Lee, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+births+after+transfer+of+rebiopsy+and+revitrification+of+blastocyst+that+had+&#8220;no+diagnosis&#8221;+following+trophectoderm+biopsy.">Live births after transfer of rebiopsy and revitrification of blastocyst that had &#8220;no diagnosis&#8221; following trophectoderm biopsy.</a> Fertility and Sterility. 106 (3), e164 (2016).</li><li>Capalbo, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+efficacy+of+blastocoel+fluid+and+spent+media+as+sources+of+DNA+for+preimplantation+genetic+testing+in+standard+clinical+conditions.">Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions.</a> Fertility and Sterility. 110 (5), 870-879 (2018).</li><li>Hammond, E. R., Shelling, A. N., Cree, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+and+mitochondrial+DNA+in+blastocoele+fluid+and+embryo+culture+medium:+evidence+and+potential+clinical+use.">Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: evidence and potential clinical use.</a> Human Reproduction. 31 (8), 1653-1661 (2016).</li><li>Vera-Rodriguez, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Origin+and+composition+of+cell-free+DNA+in+spent+medium+from+human+embryo+culture+during+preimplantation+development.">Origin and composition of cell-free DNA in spent medium from human embryo culture during preimplantation development.</a> Human Reproduction. 33 (4), 745-756 (2018).</li></ol>

只有经验丰富的熟练胚胎学家谁完成了他们的训练期应进行TE活检和胚泡性牙菌。此外,需要证人监测程序,并保证在i期间有效追踪活检胚泡从活检盘(补充图1)到活检后盘的移动(补充图1)(补充图1)),然后到化板(补充图1),最后到硝化支持(补充图1);ii) 活检TE细胞从活检盘转移到PCR管(补充图1);iii) 诊断后的加热和转移步骤。有关所有见证步骤...

现代人类胚胎学的主要目标是使每个刺激周期的活产数量最大化,并降低成本、时间和实现怀孕的努力。为了实现这一目标,应采用经验证的胚胎选择方法,在IVF周期中获得的一组胚胎中确定具有生殖能力的胚胎。根据最新证据,胚泡培养1结合全面的染色体检测和玻璃化加热的幼倍体胚胎移植(ET)是提高IVF效率的最有效框架2。显然,非倍体试验需要胚胎标本,目前主要代表从营养素(TE)中检索到的细胞,即囊胚中给予妊娠期间胚胎附件(例如胎盘)来源的部分.除了核类型分析,还可以从TE活检中评估单个基因突变,作为称为植入前基因测试的临床策略的一部分(PGT;-A用于非倍体,-SR用于结构染色体重组,-M用于单源性疾病)。其他卵母细胞/胚胎活检方法在过去几十年中已被进行解剖和临床采用,即极性身体活检和卵波粒活检。然而,现在减少了它们的使用,因为它们的程序性缺陷(例如,更高的工作量和生殖影响的风险)和诊断限制(例如单细胞分析问题)隐含地阻碍了成本、风险和好处(有关审查,请参阅3)。
在本文中,对TE活检的主要方案之一以及随后所需的保质化、加热和转移程序进行了详尽描述。此处概述的工作流非常适合繁忙的 PGT 单元。
如我们组4,5前面所述,该过程涉及完全扩张的胚泡的分子的连续打开和去除少数TE细胞(平均7-8)。与第3天激光辅助孵化为基础的胚泡活检方法6相比,这个程序可以简化IVF单元的日常时间表,其中必须及时执行诸如胚泡活检和葡萄光化等精细程序。一旦胚泡达到其完全膨胀,活检可以通过选择TE细胞进行去除,从而防止内细胞质量(ICM)的增生风险,否则会使手术具有挑战性。在文献中,也描述了胚泡活检的第三个方案,即一旦胚胎到达胚泡阶段,在手术5、7前几个小时进行激光辅助孵化。然而,这种方法更耗时,主要适合在经验有限的操作员手中实施TE活检的IVF单元,并且考虑到日工作量适中。
如果旨在进行IVF中的遗传分析,则细胞内体精子注射(ICSI)8应该是一种综合技术。同样,一个适当的培养系统,安全地将胚胎收获到胚泡阶段,对于实施TE活检策略也至关重要。足够数量的培养箱,以及使用低氧张力是这个目的的关键先决条件,不损害胚泡率9。同时,需要一个有效的冷冻保存程序来安全地管理PGT周期。在过去的十年中,牙化的实施提高了胚胎的存活率,甚至高达>99%10,11。这提供了足够的时间进行基因测试,并将胚胎移植推迟到下面的月经周期,在非刺激和可能更容易接受的子宫内膜12。
TE 活检和 Vitaita 化都是要求严格技能的任务,其有效性可能因经验不足的操作员而异。因此,在允许每个操作员临床执行这些程序之前,提倡一个特定的培训期;此外,应定期通过监测冷冻保存和活检程序的关键绩效指标(KPI)来评估操作人员技能的维持情况。每个IVF诊所应为此设置内部KPI,其中必须近似于国际财团公布的KPI和/或参考实验室公布的结果。
TE 活检、保质化加热和见证程序是我们部门验证的技术,这些技术已在之前三份出版物 11、13、14 中报告的所有操作员中标准化.

<table>
	<tbody>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cold tube rack</td>
			<td>Biocision</td>
			<td>XTPCR96</td>
			<td></td>
		</tr>
		<tr>
			<td>Electronic pipette controller</td>
			<td>Fisher Scientific</td>
			<td>710931</td>
			<td></td>
		</tr>
		<tr>
			<td>Flexipet adjustable handle set</td>
			<td>Cook</td>
			<td>G18674</td>
			<td>Stripper holder</td>
		</tr>
		<tr>
			<td>Gilson Pipetman</td>
			<td>Gilson</td>
			<td>66003</td>
			<td>p20</td>
		</tr>
		<tr>
			<td>IVF Electronic Witness System</td>
			<td>CooperSurgical Fertility &#38; Genomic Solutions</td>
			<td></td>
			<td>RI Witness ART Management System</td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Nikon</td>
			<td>Eclipse TE2000-U</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminar Flow Hood</td>
			<td>IVF TECH</td>
			<td></td>
			<td>Grade A air flow</td>
		</tr>
		<tr>
			<td>Laser objective</td>
			<td>RI</td>
			<td>Saturn 5</td>
			<td></td>
		</tr>
		<tr>
			<td>Microinjectors</td>
			<td>Nikon Narishige</td>
			<td>NT-88-V3</td>
			<td></td>
		</tr>
		<tr>
			<td>Mini centrifuge for PCR tubes</td>
			<td>Eppendorf</td>
			<td>CSLQSPIN</td>
			<td>for 0.2ml PCR tubes</td>
		</tr>
		<tr>
			<td>Stereomicroscope</td>
			<td>Leica</td>
			<td>Leica M80</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermostat</td>
			<td>Panasonic</td>
			<td>MCO-5AC-PE</td>
			<td></td>
		</tr>
		<tr>
			<td>Tri-gas incubator</td>
			<td>Panasonic</td>
			<td>MCO-5M-PE</td>
			<td>02/CO2</td>
		</tr>
		<tr>
			<td>Consumables</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy pipette</td>
			<td>RI</td>
			<td>7-71-30FB35720</td>
			<td>30&#181;m ID, flat 35&#176;C</td>
		</tr>
		<tr>
			<td>Cryolock</td>
			<td>Cryolock</td>
			<td>CL-R-CT</td>
			<td></td>
		</tr>
		<tr>
			<td>CSCM complete</td>
			<td>Irvine Scientific</td>
			<td>90165</td>
			<td>IVF culture medium supplemented with HSA</td>
		</tr>
		<tr>
			<td>Embryo Transfer Catheter</td>
			<td>Cook</td>
			<td>G17934</td>
			<td></td>
		</tr>
		<tr>
			<td>Flexipet pipette</td>
			<td>Cook</td>
			<td>G26712</td>
			<td>140&#181;m stripping pipette tip</td>
		</tr>
		<tr>
			<td>Flexipet pipette</td>
			<td>Cook</td>
			<td>G46020</td>
			<td>300&#181;m stripping pipette tips</td>
		</tr>
		<tr>
			<td>Holding pipette</td>
			<td>RI</td>
			<td>7-71-IH35/20</td>
			<td>30&#181;m ID, flat 35&#176;C</td>
		</tr>
		<tr>
			<td>Human Serum Albumin</td>
			<td>Irvine Scientific</td>
			<td>9988</td>
			<td></td>
		</tr>
		<tr>
			<td>IVF One well dish</td>
			<td>Falcon</td>
			<td>353653</td>
			<td></td>
		</tr>
		<tr>
			<td>Mineral Oil for embryo culture</td>
			<td>Irvine Scientific</td>
			<td>9305</td>
			<td></td>
		</tr>
		<tr>
			<td>Modified HTF Medium</td>
			<td>Irvine Scientific</td>
			<td>90126</td>
			<td>Hepes-Buffered medium</td>
		</tr>
		<tr>
			<td>Nuclon Delta Surface</td>
			<td>Thermofisher scientific</td>
			<td>176740</td>
			<td>IVF dish 4-well plate with sliding lid</td>
		</tr>
		<tr>
			<td>Primaria Cell culture dish</td>
			<td>Corning</td>
			<td>353802</td>
			<td>60x15mm</td>
		</tr>
		<tr>
			<td>Reproplate</td>
			<td>Kitazato</td>
			<td>83016</td>
			<td></td>
		</tr>
		<tr>
			<td>Serological pipette</td>
			<td>Falcon</td>
			<td>357551</td>
			<td>10ml</td>
		</tr>
		<tr>
			<td>Sterile disposable Gilson tips</td>
			<td>Eppendorf</td>
			<td>0030 075.021</td>
			<td>200&#181;l</td>
		</tr>
		<tr>
			<td>Tubing Kit</td>
			<td>Provided by the genetic lab</td>
			<td></td>
			<td>PCR tubes (0.2mL), loading solution, biopsy washing solution</td>
		</tr>
		<tr>
			<td>Vitrification media</td>
			<td>Kitazato</td>
			<td>VT801</td>
			<td>Equilibration and vitrification solutions</td>
		</tr>
		<tr>
			<td>Warming media</td>
			<td>Kitazato</td>
			<td>VT802</td>
			<td>Thawing and dilution solutions</td>
		</tr>
	</tbody>
</table>

human blastocyst biopsy and vitrification

在IVF周期内进行胚泡活检,通过植入前基因测试获得可靠的基因诊断。然后,理想的工作流程需要一个安全有效的牙化协议,由于诊断技术的周转时间,并在下面的自然周期中在生理子宫内膜上移植选定的胚胎。活检方法包括连续打开区域细胞和检索5-10个营养细胞(理想情况下为7-8)限制所需的操作数量和胚胎在次优环境条件下的暴露。经过适当的训练后,该技术在不同的操作者之间可重复活检的时间(±8分钟,根据每盘的胚胎数量进行活检3-22分钟),获得结论性诊断(+97.5%)和玻璃化加热的花体胚泡转移后活产率(>40%)。活检、硝化和变暖后的存活率高达99.8%。气候变暖后1.5小时的再膨胀率高达97%,主要取决于活检和抗化(理想情况下为±30分钟)、胚泡形态质量和活检日之间的时间。一般来说,最好对坍塌的胚泡进行压化;因此,在非PGT周期中,可以进行激光辅助人工收缩,以诱导胚胎在冷冻保存前崩溃。最有希望的未来观点是,在胚泡培养后对IVF培养基进行非侵入性分析,作为胚胎DNA的假定来源。然而,这种潜在的前卫仍在调查之中,还需要定义和验证一个可靠的协议。

图 6显示了活检程序的所有结果方案,可用于标准化协议并监控每个操作员的性能。主要程序结果是完成活检/活检的时间;主要技术结果是基因测试后产生的图的质量,可能导致结论性或无结论性诊断,后者要求对未诊断的胚泡进行重新活检;主要生物结果是变暖后的低温生存率和再膨胀率与退化率;最后,主要临床结果为玻璃化加热胚泡转移后的活产率。在...

Watch this Scientific Journal Video about Human Blastocyst Biopsy and Vitrification at JoVE.com

Human Blastocyst Biopsy and Vitrification

需要进行胚泡活检和保并口授,以便有效地进行植入前基因检测。一种方法,导致在授精后的第5-7天连续打开区域细胞和检索7-8个营养细胞,限制了所需的操作次数和胚胎在次优环境条件下的暴露。

人类囊胚活检和细胞化

Blastocyst biopsy is performed to obtain a reliable genetic diagnosis during IVF cycles with preimplantation genetic testing. Then, the ideal workflow ...

爆细胞活检和玻璃化都是试管婴儿的一场革命，体外受精使来自世界各地的胚胎学家能够对人类胚胎进行植入前基因测试，而胚胎风险最小。Trophectoderm活检使从完全生长的胚胎中检索到大约7个细胞，从而能够进行可靠的下游基因分析。此外，迄今为止，尚未显示对这种做法的影响。此工作流程的临床效率使单基因条件的患者和所有在其囊肿内染色体异常风险增加的患者受益于植入前基因测试。目前，胚胎选择策略仍有改进的余地。例如，miRnomic 和蛋白质组分析代表有趣的选项，以补充单次营养异常活检的植入前基因测试。缺乏经验的操作员经常难以打开和穿透佐纳佩鲁西达。这只是一个焦点问题。爆震、激光目标和移液器应在同一焦点平面上。在用患者的详细信息标记活检盘后，用永久的、无毒的标记，用胚胎和周期 ID 将每 10 微升的 HEPES 缓冲介质数数。 使用 300 微米剥离移液器，将可行、完全展开的爆震转移到活检盘的第一滴，并冲洗滴以去除多余的培养基，然后再将胚胎转移到第二滴。将盘子放在倒置显微镜下，用活检盘第三滴的介质对活检移液器进行加热。在20倍的放大倍率下，定向胚胎，在7点钟时获得内细胞质量的清晰视图，并固定胚胎在保持移液器上。专注于佐纳佩普鲁西达，使移液器和爆震都在同一焦点平面上。切换到激光目标，将激光笔定位在内细胞质量的对面。用两到三个激光脉冲钻取佐纳佩普利达，然后轻轻地将活检移液器压在佐纳佩普鲁西达上，使介质通过突破口吹出，将营养细胞从内部表面分离出来。当营养细胞分离时，通过孔进入，用温和的吸力将七到八个营养细胞吸入活检移液器。在应用适度吸力拉伸目标细胞时，稍微向后移动活检移液器，将激光引导至吸气细胞的最薄部分。在细胞之间的交界处发射两到五个激光脉冲，将目标细胞与胚胎体分离。当营养球菌片段与爆震分离时，将片段释放到远离囊肿的同一活检滴中，以防止片段被吸回活检移液器中。从保持移液器中释放爆震，并及时升起两个移液器，以防止碎片粘在移液器上。然后为质量控制目的对活检片段进行成像。当所有胚胎都像刚刚演示的一样进行活检时，将活检盘移回层流罩，并标记后活检培养皿与夫妇 ID 和每个滴与胚胎和循环 ID。当有目击证人在场的情况下，用干净的IVF介质冲洗胚胎，将胚胎移动到其在活检后盘中的相应下降。然后将活检后盘放在37摄氏度、6%的二氧化碳和5%的氧气培养箱中。在证人在场的情况下，在室温下在层流罩内，用永久的、无毒的标记标记 PCR 管。用活检胚胎 ID 标记 60 x 15 毫米管培养皿的盖子，并在该盘中加入两滴 10 微升活检洗涤溶液。在立体显微镜下，用活检洗涤溶液从管盘的第二滴中，用活检洗涤液来放置 140 微米脱液器，以可视化营养球状片段。轻轻释放一些活检洗涤溶液，在营养球片碎片上，将碎片加载到剥离移液器中。将碎片移动到活检洗涤溶液的第二滴，并小心冲洗组织两到三次。使用装载溶液将营养球状碎片转移到贴有适当标记的 PCR 管的底部，注意避免用剥离移液器的尖端接触管壁。当所有碎片都添加到它们的管子中时，在一个小型离心机中旋转所有管子几秒钟，以沉淀碎片。然后将样品储存在零下20摄氏度，直到被运到推荐基因实验室进行测试。在30分钟内的营养细胞活检，标记一个玻璃化板与病人的详细信息和必须玻璃化的囊肿的ID。使用特殊的耐低温冷冻棒，用患者的姓名、夫妻身份证、将加载到其上的胚胎的 ID 以及手术日期来标记玻璃化支架。在室温下，为每种将进行玻璃化的气囊分配 300 微升的平衡溶液，并在有目击证人在场的情况下，使用 300 微米剥离移液器将爆炸囊移入一卷平衡溶液中。将防爆液留在平衡溶液中 13 至 15 分钟。在卷的初始收缩后，将观察到逐渐的重新膨胀。在平衡结束时，在玻璃化板的第二孔中加入300微升的玻璃化溶液，并将完全重新膨胀的爆炸囊液转移到玻璃化溶液中一分钟。在孵化结束时，冲洗囊肿以稀释平衡溶液，并在有目击者在场的情况下，将爆炸囊加载到玻璃化支撑上。去除多余的玻璃化溶液。一个微妙的溶液膜应该围绕爆炸。将玻璃化支架插入液氮中，并大力移动支架，以降低靠近试样形成气泡的风险。然后，当玻璃化支撑仍浸入液氮中时，将保护盖放在支撑上。在这一具有代表性的两年期间进行的1 544次营养细胞活检程序中，每个手术进行1至4次爆震活检，每个程序3至22分钟。在这些图中，显示了每个操作员在研究三个月时进行活检的均值，平均总值为 8.24 分钟。获取每个活检片段的图像以用于质量控制，以评估无定论诊断的原因是否与片段的尺寸和/或质量、管材或遗传实验室中样品处理中的一些问题有关。在这项研究中，572个欧倍体胚胎在诊断出后被加热接受胚胎移植。所有囊肿在30分钟内全部发生，2.4%的囊肿在31至90分钟内不重新膨胀，4.9%的囊肿在重新温化后90分钟以上不重新膨胀。特别是对于质量差和/或第七天爆震，活检和玻璃化之间的时间似乎至关重要，以实现变暖后重新膨胀。在暴露营养细胞和发射之间的交界处时，在打开 zona pellucida 时注意聚焦，并注意防止在玻璃化之前重新膨胀囊肿。另外两种胚胎活检方法也存在，两者都需要佐纳·佩鲁西达的开口和等待营养细胞的自发拱形。这些方法比较容易，但需要更多的操作，而且更耗时。该工作流程的功效允许在IVF中实施分子技术，以尝试在胚胎移植前选择植入机率最高的胚胎。请记住，不要使用任何可能对胚胎有毒的装置或溶液，并使用无DNA材料防止DNA污染。

Research

JoVE Journal

Developmental Biology

Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle

从人类骨骼肌成肌原细胞和成纤维细胞的分离和定量免疫细胞化学表征

The repair and regeneration of skeletal muscle requires the action of satellite cells, which are the resident muscle stem cells. These can be isolated ...

科研

发育生物学

Three-dimensional Quantification of Dendritic Spines from Pyramidal Neurons Derived from Human Induced Pluripotent Stem Cells

树突棘从锥体神经元的三维定量从人类诱导多能干细胞衍生

Dendritic spines are small protrusions that correspond to the post-synaptic compartments of excitatory synapses in the central nervous system. They are ...

Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research

培养人多能性和神经干细胞的基本和临床前研究一个封闭的细胞培养系统

This paper describes how to use a custom manufactured, commercially available enclosed cell culture system for basic and preclinical research. Biosafety ...

Modified MicroSecure Vitrification: A Safe, Simple and Highly Effective Cryopreservation Procedure for Human Blastocysts

修改MicroSecure玻璃化：一种安全，简便，高效的冷冻程序人胚泡

Clinical embryo vitrification evolved with the development of unique vitrification devices in the 21st century and with the misconception that ultra-rapid ...

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

细胞聚集法评价果蝇切口与反式配体的结合及其对顺式配体的抑制作用

Notch signaling is an evolutionarily conserved cell-cell communication system used broadly in animal development and adult maintenance. Interaction of the ...

Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model

家兔模型最佳胚胎阶段微创胚胎移植与胚胎玻璃化

Assisted reproductive techniques (ARTs), such as in vitro embryo culture or embryo cryopreservation, affect natural development patterns with perinatal ...

Isolation, Culture, Characterization, and Differentiation of Human Muscle Progenitor Cells from the Skeletal Muscle Biopsy Procedure

从骨骼肌活检程序分离、培养、表征和分化人类肌肉祖细胞

The use of primary human tissue and cells is ideal for the investigation of biological and physiological processes such as the skeletal muscle ...

Determining Bile Duct Density in the Mouse Liver

确定小鼠肝脏中的胆汁密度

Mouse is broadly used as a model organism to study biliary diseases. To evaluate the development and function of the biliary system, various techniques ...

Human Egg Maturity Assessment and Its Clinical Application

人卵成熟度评估及其临床应用

The optimal timing of intracytoplasmic sperm injection (ICSI) is of a serious concern for fertility programs because untimely sperm entry diminishes the ...

Generation of Multicellular Human Primary Endometrial Organoids

多细胞人类原发性子宫内膜器官的生成

The human endometrium is one of the most hormonally responsive tissues in the body and is essential for the establishment of pregnancy. This tissue can ...