Name: co-transplantation of human ovarian tissue with engineered endothelial cells: a cell-based strategy combining accelerated perfusion with direct paracrine delivery
Uploaded: 2018-05-16T14:00:00
Description: Watch this Scientific Journal Video about Co-transplantation of Human Ovarian Tissue with Engineered Endothelial Cells: A Cell-based Strategy Combining Accelerated Perfusion with Direct Paracrine Deli

奥马尔. 亚历山大的人为例证。 L.M. 获得了康奈尔临床和翻译科学中心的试验奖和 ASRM 研究补助金的支持。 作者感谢詹姆斯实验室成员对手稿的批判性阅读。

所有涉及动物的程序都已由威尔康奈尔医学院的机构动物护理和使用委员会 (IACUC) 批准。所有使用卵巢组织的异种移植实验均按照相关的指导原则和规定进行。人卵巢组织收集的病人计划化疗或放疗的癌症治疗或前骨髓移植。威尔康奈尔医学院的机构审查委员会批准了为潜在自体使用的组织的收集, 并在患者知情同意下, 捐赠多达10% 的卵巢组织供研究使用。
1. 人类卵巢组织的?...

<ol>
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K., 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=Outcomes+of+transplantations+of+cryopreserved+ovarian+tissue+to+41+women+in+Denmark.">Outcomes of transplantations of cryopreserved ovarian tissue to 41 women in Denmark.</a> Hum Reprod. 30 (12), 2838-2845 (2015).</li><li>Oktay, K., Newton, H., Aubard, Y., Salha, O., Gosden, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cryopreservation+of+immature+human+oocytes+and+ovarian+tissue:+an+emerging+technology?.">Cryopreservation of immature human oocytes and ovarian tissue: an emerging technology?.</a> Fertil Steril. 69 (1), 1-7 (1998).</li><li>Shultz, L. 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=Human+lymphoid+and+myeloid+cell+development+in+NOD/LtSz-scid+IL2R+gamma+null+mice+engrafted+with+mobilized+human+hemopoietic+stem+cells.">Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells.</a> J Immunol. 174 (10), 6477-6489 (2005).</li><li>Ramalingam, R., Rafii, S., Worgall, S., Brough, D. E., Crystal, R. 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Potential mechanisms of ovarian injury.</a> Hum Reprod. 22 (6), 1626-1633 (2007).</li><li>Assidi, 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=Identification+of+potential+markers+of+oocyte+competence+expressed+in+bovine+cumulus+cells+matured+with+follicle-stimulating+hormone+and/or+phorbol+myristate+acetate+in+vitro.">Identification of potential markers of oocyte competence expressed in bovine cumulus cells matured with follicle-stimulating hormone and/or phorbol myristate acetate in vitro.</a> Biol Reprod. 79 (2), 209-222 (2008).</li><li>Thakur, S. C., Datta, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Higher+expression+of+hyaluronan+binding+protein+1+(HABP1/p32/gC1qR/SF2)+during+follicular+development+and+cumulus+oocyte+complex+maturation+in+rat.">Higher expression of hyaluronan binding protein 1 (HABP1/p32/gC1qR/SF2) during follicular development and cumulus oocyte complex maturation in rat.</a> Mol Reprod Dev. 75 (3), 429-438 (2008).</li><li>Dolmans, M. 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=Short-term+transplantation+of+isolated+human+ovarian+follicles+and+cortical+tissue+into+nude+mice.">Short-term transplantation of isolated human ovarian follicles and cortical tissue into nude mice.</a> Reproduction. 134 (2), 253-262 (2007).</li><li>Amorim, C. 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=Impact+of+freezing+and+thawing+of+human+ovarian+tissue+on+follicular+growth+after+long-term+xenotransplantation.">Impact of freezing and thawing of human ovarian tissue on follicular growth after long-term xenotransplantation.</a> J Assist Reprod Genet. 28 (12), 1157-1165 (2011).</li><li>Kawamura, K., 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=Hippo+signaling+disruption+and+Akt+stimulation+of+ovarian+follicles+for+infertility+treatment.">Hippo signaling disruption and Akt stimulation of ovarian follicles for infertility treatment.</a> Proc Natl Acad Sci U S A. 110 (43), 17474-17479 (2013).</li><li>Suzuki, N., 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=Successful+fertility+preservation+following+ovarian+tissue+vitrification+in+patients+with+primary+ovarian+insufficiency.">Successful fertility preservation following ovarian tissue vitrification in patients with primary ovarian insufficiency.</a> Hum Reprod. 30 (3), 608-615 (2015).</li><li>Campbell, B. K., Clinton, M., Webb, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+anti-M&#252;llerian+hormone+(AMH)+during+follicle+development+in+a+monovulatory+species+(sheep).">The role of anti-M&#252;llerian hormone (AMH) during follicle development in a monovulatory species (sheep).</a> Endocrinology. 153 (9), 4533-4543 (2012).</li><li>Donnez, J., 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=Restoration+of+ovarian+activity+and+pregnancy+after+transplantation+of+cryopreserved+ovarian+tissue:+a+review+of+60+cases+of+reimplantation.">Restoration of ovarian activity and pregnancy after transplantation of cryopreserved ovarian tissue: a review of 60 cases of reimplantation.</a> Fertil Steril. 99 (6), 1503-1513 (2013).</li><li>Ferreira, 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=The+effects+of+sample+size+on+the+outcome+of+ovarian+tissue+cryopreservation.">The effects of sample size on the outcome of ovarian tissue cryopreservation.</a> Reprod Domest Anim. 45 (1), 99-102 (2010).</li><li>Gavish, Z., Peer, G., Roness, H., Cohen, Y., Meirow, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Follicle+activation+and+'burn-out'+contribute+to+post-transplantation+follicle+loss+in+ovarian+tissue+grafts:+the+effect+of+graft+thickness.">Follicle activation and 'burn-out' contribute to post-transplantation follicle loss in ovarian tissue grafts: the effect of graft thickness.</a> Hum Reprod. 30 (4), 1003 (2015).</li><li>Donnez, J., Dolmans, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fertility+Preservation+in+Women.">Fertility Preservation in Women.</a> N Engl J Med. 377 (17), 1657-1665 (2017).</li><li>Salama, M., Woodruff, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+advances+in+ovarian+autotransplantation+to+restore+fertility+in+cancer+patients.">New advances in ovarian autotransplantation to restore fertility in cancer patients.</a> Cancer Metastasis Rev. 34 (4), 807-822 (2015).</li><li>Donnez, J., Dolmans, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+cortex+transplantation:+60+reported+live+births+brings+the+success+and+worldwide+expansion+of+the+technique+towards+routine+clinical+practice.">Ovarian cortex transplantation: 60 reported live births brings the success and worldwide expansion of the technique towards routine clinical practice.</a> J Assist Reprod Genet. 32 (8), 1167-1170 (2015).</li><li>Meirow, 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=Transplantations+of+frozen-thawed+ovarian+tissue+demonstrate+high+reproductive+performance+and+the+need+to+revise+restrictive+criteria.">Transplantations of frozen-thawed ovarian tissue demonstrate high reproductive performance and the need to revise restrictive criteria.</a> Fertil Steril. 106 (2), 467-474 (2016).</li><li>Kalich-Philosoph, L., 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=Cyclophosphamide+triggers+follicle+activation+and+"burnout";+AS101+prevents+follicle+loss+and+preserves+fertility.">Cyclophosphamide triggers follicle activation and "burnout"; AS101 prevents follicle loss and preserves fertility.</a> Sci Transl Med. 5 (185), 185ra162 (2013).</li><li>Kano, 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=AMH/MIS+as+a+contraceptive+that+protects+the+ovarian+reserve+during+chemotherapy.">AMH/MIS as a contraceptive that protects the ovarian reserve during chemotherapy.</a> Proc Natl Acad Sci U S A. 114 (9), E1688-E1697 (2017).</li></ol>

在这里我们证明, 联合移植的执行官提供了一个重要的好处, 卵巢组织的生存能力和功能后, 移植的小鼠。未设置卵巢组织自动移植的临床应用标准和最佳参数 (大小, 移植部位, 移植时间,等)。32,33,34对于滤泡池的增强恢复仍未定义。当进行自动移植时, 解冻皮层卵巢组织的缺血性移植主要表现在骨盆部位, 如其余的卵巢、?...

癌症仍然是发达国家死亡的主要原因之一, 但数十年的研究已经取得了重大进展, 在大多数类型的癌症, 并在某些情况下, 几乎加倍的生存率1。不幸的是, 化疗药物经常 gonadotoxic, 耗尽卵巢原始卵泡的储备, 并减少生育率2。这种日益增长的人口可以受益于多种生育方法, 包括卵母细胞和/或胚胎超低温保存, 但是, 需要迅速启动癌症治疗和青春期前患者的患者不符合这些选择。另外, 一些患者选择在进行治疗方案之前 cryopreserve 卵巢组织, 并在恢复和缓解后, 自动移植组织恢复生育率3。然而, 到目前为止, 自动移植后的移植生存和卵泡输出仍然相对较低4, 主要是由于组织缺血和缺氧5,6,7。尽管许多努力改善卵巢皮质移植的生存能力使用抗氧化剂8,9, 支持血管生成素10,11,12,1 3或机械操作14, 5 至7天窗口移植后的移植物缺血损害了移植7的生存和生存。为了解决这一问题, 我们制定了一个基于细胞的策略, 以促进宿主和移植血管的吻合, 从而加速卵巢组织的再灌注。
除了在移植后的窗口中对接枝卵巢组织的缺血性侮辱外, 卵泡间信号的中断可能导致池的耗尽15,16。由于外源性内皮细胞 (执行官) 有助于在移植周围的稳定和功能的血管, 他们提供了一个独特的机会, 向移植组织传达定义的分子输入。作为一项原则的证明, 执行官们被设计来表达超生理水平的抗苗勒激素 (AMH), 一个成员的转化生长因子 beta (TGFβ) 超家族, 已被证明限制卵泡生长17。用控制和 AMH 表达细胞联合移植移植滤泡分布的比较验证了工程主管的生物活性和效力。
总之, 通过提高移植的生存能力和抑制卵泡池的过早动员, 这种方法可以提高在生育保护患者中自动移植卵巢组织的生产率。此外, 基于 ExEC 的平台还能对与卵泡发育有关的分子调控器进行实验性审讯。

<table>
	<tbody>
		<tr>
			<td>Leibovitz&#8217;s L-15 medium</td>
			<td>Gibco</td>
			<td>11415064</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic</td>
			<td>Gibco</td>
			<td>15240062</td>
			<td>Anti-Anti X100</td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma</td>
			<td>S 1888</td>
			<td></td>
		</tr>
		<tr>
			<td>Fibrinogen</td>
			<td>Sigma</td>
			<td>F 8630</td>
			<td>from bovine plasma</td>
		</tr>
		<tr>
			<td>Thrombin</td>
			<td>Sigma</td>
			<td>T 1063</td>
			<td>from human plasma</td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma</td>
			<td>D 2650</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Gibco</td>
			<td>12491015</td>
			<td></td>
		</tr>
		<tr>
			<td>Enzyme Cell Detachment Medium</td>
			<td>Invitrogen</td>
			<td>00-4555-56</td>
			<td>Accutase</td>
		</tr>
		<tr>
			<td>Plastic paraffin film</td>
			<td>Bemis NA</td>
			<td></td>
			<td>Parafilm M</td>
		</tr>
		<tr>
			<td>Surgical paper tape 2.5 cm</td>
			<td>3M</td>
			<td>1530-1</td>
			<td>Micropore</td>
		</tr>
		<tr>
			<td>Surgical Paper tape 1.25 cm</td>
			<td>3M</td>
			<td>1530-0</td>
			<td>Micropore</td>
		</tr>
		<tr>
			<td>Perforated plastic Surgical tape 1.25 cm</td>
			<td>3M</td>
			<td>1527-0</td>
			<td>Transpore</td>
		</tr>
		<tr>
			<td>Monofilament Absorbable Suture</td>
			<td>Covidien</td>
			<td>UM-203</td>
			<td>Biosyn</td>
		</tr>
		<tr>
			<td>Braided Absorbable Suture</td>
			<td>Covidien</td>
			<td>GL-889</td>
			<td>Polysorb</td>
		</tr>
		<tr>
			<td>Povidone-iodine Solution USP 10%</td>
			<td>Purdue Products</td>
			<td>67618-153-01</td>
			<td>Betadine Solution Swab Stick</td>
		</tr>
		<tr>
			<td>Cryoviales</td>
			<td>Nunc</td>
			<td>377267</td>
			<td>CryoTube</td>
		</tr>
		<tr>
			<td>sterile ocular lubricant</td>
			<td>Dechra</td>
			<td>17033-211-38</td>
			<td>Puralube</td>
		</tr>
		<tr>
			<td>1.7 ml micro-centrifuge tube</td>
			<td>Denville</td>
			<td>C-2172</td>
			<td>Eppendorf</td>
		</tr>
		<tr>
			<td>Anasthesia system</td>
			<td>VetEquip</td>
			<td>V-1 table top system with scavenging</td>
			<td></td>
		</tr>
		<tr>
			<td>Endothelial cells</td>
			<td>Angiocrine Biosciences, Inc., San Diego, CA, USA</td>
			<td></td>
			<td>Isolated, transfected with E4-ORF- 1 and labeled endothelial cells</td>
		</tr>
		<tr>
			<td>Trichrome stain</td>
			<td>Sigma</td>
			<td>HT15-1kt</td>
			<td>Trichrome Stain (Masson) Kit</td>
		</tr>
		<tr>
			<td>Isolectin</td>
			<td>Invitrogen</td>
			<td>I32450</td>
			<td>isolectin GS-IB4 From Griffonia simplicifolia, Alexa Fluor&#8482; 647 Conjugate</td>
		</tr>
	</tbody>
</table>

co-transplantation of human ovarian tissue with engineered endothelial cells: a cell-based strategy combining accelerated perfusion with direct paracrine delivery

不孕是化疗和/或放疗的经常性副作用, 对某些患者来说, 冷冻的卵母细胞或胚胎不是一种选择。作为替代, 越来越多的这些患者选择 cryopreserve 卵巢组织的移植后恢复和缓解。尽管保存的卵巢组织自动移植患者的预后有所改善, 但移植组织的有效血管重建仍然是一个主要障碍。为了减轻缺血, 从而改善自动移植患者的预后, 我们开发了一种以血管细胞为基础的加速卵巢组织灌注的策略。本文描述了一种在小鼠同种异体模型中联合移植保存卵巢组织的外源性内皮细胞 (执行官) 的方法。我们扩大这一做法, 雇用已设计为组成性表达抗苗勒激素 (AMH) 的执行官, 从而使持续分泌信号输入到卵巢移植。联合移植与执行官增加卵泡体积和改善窦卵泡发育, 和 AMH 表达的执行官促进保留静态原始卵泡。这一联合战略可能是一个有用的工具, 以减轻缺血和调节卵泡激活的背景下的生育率保存和/或不育的大。

为了确定联合移植的执行官是否为患者的组织提供了好处, 解冻后的卵巢皮质带被分成同等大小的部分, 并嫁接为免疫性损害, 点头, 伽玛 (核供应), 小鼠。在单独嵌入纤维蛋白血块 (无 ECs) 和其他包含执行官 (图 1a) 的一侧, 每只鼠标都充当自己的控制项。执行官是通过从人脐带分离的原发性内皮和随后的治疗与腺病毒基因片段 E4-ORF1, 如前所述<sup class="...

Watch this Scientific Journal Video about Co-transplantation of Human Ovarian Tissue with Engineered Endothelial Cells: A Cell-based Strategy Combining Accelerated Perfusion with Direct Paracrine Deli

Co-transplantation of Human Ovarian Tissue with Engineered Endothelial Cells: A Cell-based Strategy Combining Accelerated Perfusion with Direct Paracrine Delivery

对某些患者来说, 保留生育能力的唯一选择是保存卵巢组织。不幸的是, 延迟血管重建破坏卵泡活力。在这里, 我们提出一个协议, 共同移植人卵巢组织与内皮细胞作为一种基于细胞的战略结合加速灌注与直接分泌交付的生物活性分子。

人卵巢组织与工程化内皮细胞联合移植: 一种以细胞为基础的快速灌注与直接分泌的融合策略

Infertility is a frequent side effect of chemotherapy and/or radiotherapy and for some patients, cryopreservation of oocytes or embryos is not an option. ...

The overall goal of this protocol is to xenograft cryopreserved human ovarian tissue in combination with engineered endothelial cells that accelerate re-perfusion and provide paracrine stimulus that modulates follicular activation within the graft. This method provides a cell-based means of improving post-transplant viability of cryopreserved ovarian tissue and more broadly, establishes a robust model system for interrogating molecular regulation of human folliculogenesis. The main advantage of this technique is that it uses a single vehicle that of endothelial cells to both mitigate ischemia and provide a sustained source of secreted factors to engraft the tissue.So this method provide global benefit for tissue viability. It can also provide insight into molecular regulators that have been implicated in follicular development. To make a fibrin clot, work quickly and add 360 microliters of HEPES buffer to 200 microliters of the fibrinogen stock solution.Gently pipette to mix and keep the tube on ice. In the second microfuge tube, prepare thrombin working solution by adding 148.7 microliters of DMEM to 50 microliters of thrombin stock solution. Gently pipette to mix.Then add 1.3 microliters of one mole per liter of calcium chloride. Gently pipette to mix and keep the sample on ice. Using the enzyme cell detachment medium Accutase, detach previously prepared engineered endothelial cells from the plate and transfer them to a third microfuge tube.Then, spin down the cells. Next, using a hemocytometer and a covered glass, count the cells. To calculate the number of cells needed for the engraftment, convert a 20, 000 times area of ovarian tissue into millimeter squared by using a ruler to measure the dimensions.The multiply the area by 20, 000 to determine the number of cells needed for the engraftment. Spin down the required cells. And use DMEM to re-suspend the pellet for a total volume of 16.8 microliters.Keep the cell suspension on ice. Next, place a piece of ovarian tissue on a sterile gauze sponge for a few seconds to dry it. Then transfer the tissue onto a previously prepared plastic paraffin film in a 50 millimeter petri dish.Add 39.2 microliters of the fibrinogen solution into the prepared cell suspension. Mix by gentle pipetting, and keep the suspension on ice. Then working quickly, add 14 microliters of the thrombin solution to the tube and gently pipette once to mix.Pipette the mixture on top of the ovarian tissue in the shape of an elongated droplet. Incubate the petri dish with the clot at 37 degrees Celsius. Work quickly once you have added the thrombin solution to the fibrinogen cell suspension.Synchronizing the future preparation in surgery is a challenge and it may be prudent to perform a practice run prior to expending precious human tissue. After anesthetizing the 10 to 14 week old female NSG mouse, and preparing it for surgery according to the text protocol, heat the animal and monitor the body temperature throughout the procedure. With a povidone iodine swab stick, clean the trimmed area and then use sterile alcohol to wipe it off.Next, place a drop of sterile ocular lubricant vet ointment over each of the mouse's eyes to protect them from dehydration and damage to the cornea. After performing a bilateral oophorectomy according to the text protocol, inject buprenorphine subcutaneous. Then make a horizontal incision in the fascia above the gluteus maximus at a length that fits the clot dimensions.Create a pocket within this virtual space by opening the scissors underneath the fascia. Pick up one piece of previously prepared encapsulated cortical tissue and place it in the pocket. Then use six O braided absorbable suture to suture the fascia before repeating the co-transplant on the contra-lateral side.Close the dorsal wall with four O monofilament absorbable suture in simple interrupted stitches. Inject lidocaine subcutaneous at the incision site. Then use a sterile alcohol prep to clean the skin and carry out post-operative care according to the text protocol.As seen here, two weeks after bilateral engraftment of E4-ORF1 treated human umbilical vein endothelial cells into NSG female mice, functionally perfused vessels formed from GFP labeled exogenous ECs or ExECs at the interface of host tissue and the graft. ExECs formed functional vessels when co-transplanted with ovarian tissue in cortical pieces co-transplanted with the mouse or human ExECs, increase the number of surviving follicles relative to control grafts. This increase in follicle development was associated with a decreased fibrotic area in ExEC assisted grafts.Final fibrotic area values were calculated for each graft as the average of three sections analyzed. Importantly, grafts that were co-transplanted with human foreskin fibroblasts showed poor tissue viability and relatively few follicles. 14 weeks after engraftment, mice were stimulated with menotropins.And the progression of follicle growth was monitored via MRI. Developing follicles were noted in both control and ExEC assisted grafts. However, more and larger sized follicles were noticed in the ExEC assisted grafts.Compared to antral follicles derived from the same patient's tissue, the granulosa cell layer in the more advanced follicle displayed increased expression of the ovulatory markers CD44 and HABP1, as well as increased cell death and reduced proliferation. After watching this video, you will understand how to perform co-transplantation of cryopreserved ovarian tissue with endothelial cells in a fibrin clot. And once mastered, you should be able to complete this protocol in less than one hour.Building on this approach, endothelial cells can be engineered to express any combination of secreted factors, thereby enabling interrogation of signaling pathways that may be relevant to fertility preservation, folliculogenesis, and/or tissue transplantation.

Research

JoVE Journal

Bioengineering

Sensing of Barrier Tissue Disruption with an Organic Electrochemical Transistor

阻隔组织破坏与有机电化学晶体管传感

The gastrointestinal tract is an example of barrier tissue that provides a physical barrier against entry of pathogens and toxins, while allowing the ...

科研

生物工程

Tissue Engineering of Tumor Stromal Microenvironment with Application to Cancer Cell Invasion

肿瘤微环境基质与应用癌细胞侵袭组织工程

3D organotypic cultures of epithelial cells on a matrix embedded with mesenchymal cells are widely used to study epithelial cell differentiation and ...

Generation and Grafting of Tissue-engineered Vessels in a Mouse Model

组织工程血管在小鼠模型的生成和嫁接

The construction of vascular conduits is a fundamental strategy for surgical repair of damaged and injured vessels resulting from cardiovascular diseases. ...

Production, Characterization and Potential Uses of a 3D Tissue-engineered Human Esophageal Mucosal Model

三维组织工程人食管黏膜型号生产，表征和潜在用途

The incidence of both esophageal adenocarcinoma and its precursor, Barrett&#8217;s Metaplasia, are rising rapidly in the western world. Furthermore ...

Delivery of Proteins, Peptides or Cell-impermeable Small Molecules into Live Cells by Incubation with the Endosomolytic Reagent dfTAT

交货蛋白质，多肽或细胞不可渗透的小分子进入活细胞被孵化与核内体溶解试剂dfTAT

Macromolecular delivery strategies typically utilize the endocytic pathway as a route of cellular entry. However, endosomal entrapment severely limits the ...

Automated Contraction Analysis of Human Engineered Heart Tissue for Cardiac Drug Safety Screening

人工程心脏组织的收缩自动分析心脏药物安全筛选

Cardiac tissue engineering describes techniques to constitute three dimensional force-generating engineered tissues. For the implementation of these ...

Microdissection of Primary Renal Tissue Segments and Incorporation with Novel Scaffold-free Construct Technology

原发性肾组织段的显微解剖与新型无支架结构技术的结合

Kidney transplantation is now a mainstream therapy for end-stage renal disease. However, with approximately 96,000 people on the waiting list and only ...

A Novel Tenorrhaphy Suture Technique with Tissue Engineered Collagen Graft to Repair Large Tendon Defects

一种新型的腱绦缝合技术，采用组织工程胶原蛋白移植物修复大肌腱缺损

Surgical management of large tendon defects with tendon grafts is challenging, as there are a finite number of sites where donors can be readily ...

Three-dimensional Patterning of Engineered Biofilms with a Do-it-yourself Bioprinter

工程生物膜的三维造型

Biofilms are aggregates of bacteria embedded in a self-produced spatially-patterned extracellular matrix. Bacteria within a biofilm develop enhanced ...

Establishing Single-Cell Based Co-Cultures in a Deterministic Manner with a Microfluidic Chip

使用微流体芯片以确定性方式建立基于单细胞的共培养物

Cell co-culture assays have been widely used for studying cell-cell interactions between different cell types to better understand the biology of diseases ...