<meta charset="utf8"></head><body>ビトリファイドヒト卵巣皮質組織における卵胞生存率評価

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1.8 mL vials</td>
			<td>VWR International GmbH</td>
			<td>479-6837</td>
			<td></td>
		</tr>
		<tr>
			<td>10 mL serological pipette</td>
			<td>Sarstedt</td>
			<td>86.1254.001</td>
			<td></td>
		</tr>
		<tr>
			<td>4 well plate</td>
			<td>Gynemed</td>
			<td>GYOOPW-FW04</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL Tube</td>
			<td>Sarstedt</td>
			<td>62.559.001</td>
			<td></td>
		</tr>
		<tr>
			<td>6 well plates</td>
			<td>Sarstedt</td>
			<td>83.3920</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacillol AF</td>
			<td>Hartmann</td>
			<td>973385</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcein AM</td>
			<td>Merck</td>
			<td>17783</td>
			<td></td>
		</tr>
		<tr>
			<td>Collagenase type 1A</td>
			<td>Merck</td>
			<td>C2674</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryosure DMSO</td>
			<td>WAK Chemie</td>
			<td>WAK-DMSO-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Custodiol</td>
			<td>Dr. Franz K&#246;hler Chemie</td>
			<td>00867288</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS CTS</td>
			<td>Gibco Life technologies</td>
			<td>A12856-01</td>
			<td></td>
		</tr>
		<tr>
			<td>ErgoOne pipette aid</td>
			<td>Starlab</td>
			<td>S7166-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylene glycol</td>
			<td>Sigma Aldrich</td>
			<td>102466</td>
			<td></td>
		</tr>
		<tr>
			<td>Euronda sterilization container</td>
			<td>euronda</td>
			<td>282021</td>
			<td></td>
		</tr>
		<tr>
			<td>G-MOPS+</td>
			<td>Vitrolife</td>
			<td>10130</td>
			<td></td>
		</tr>
		<tr>
			<td>Metal meshes</td>
			<td>Sigma Aldrich</td>
			<td>S0770</td>
			<td></td>
		</tr>
		<tr>
			<td>Metzenbaum scissors</td>
			<td>world precision instruments</td>
			<td>501262102</td>
			<td></td>
		</tr>
		<tr>
			<td>N-Bath System</td>
			<td>Nterilizer</td>
			<td>N-Bath 3.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyvinylpyrrolidone (PVP)</td>
			<td>SAGE</td>
			<td>ART-4005</td>
			<td></td>
		</tr>
		<tr>
			<td>Serum substitute supplement (SSS)</td>
			<td>Fujifilm Irvine scientific</td>
			<td>99193</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile cup</td>
			<td>Sarstedt</td>
			<td>75.562.105</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile forceps</td>
			<td>Carl Roth</td>
			<td>KL05.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Merck</td>
			<td>S0389</td>
			<td></td>
		</tr>
	</tbody>
</table>

vitrification of ovarian cortex tissue to achieve a glassy state of aggregation

Cryopreservation of ovarian tissue is an important option for fertility preservation. Explanted tissue containing ovarian follicles, in which oocytes are embedded, is cryopreserved. After storage, the ovarian tissue can be thawed, warmed, and reimplanted in the patient. For viable cells or tissue, two cryopreservation methods are available: slow freezing and vitrification1.
Vitrification is used to preserve biological materials, such as embryos and oocytes, with superior survival rates compared to the slow freezing protocol1,2,3,4. Slow freezing has limitations, such as ice crystal formation, which can potentially damage cell and tissue structures. However, slow freezing is an important cryopreservation approach that facilitates the long-term storage of biological samples, and the functionality of this method has been widely proven5. Vitrification induces a glassy state of aggregation, preventing ice crystal formation6,7. On a technical level, vitrification significantly simplifies the cryopreservation procedure by reducing equipment maintenance, decreasing the likelihood of technical errors, and shortening the duration of the cryopreservation process8,9. In female fertility preservation, ovarian tissue cryopreservation is a decisive approach prior to cancer treatment10. Different groups have successfully demonstrated the concept of cryopreservation, thawing, and transplantation of tissue based on the slow freezing protocol11,12,13,14, which is currently regarded as the standard approach15.
Vitrification of ovarian tissue is regarded as a promising alternative method16,17,18,19,20,21, in terms of resource-saving22, follicular survival rates, DNA fragmentation levels, and balanced angiogenic potential23,24,25,26,27. This is substantiated by successful deliveries in Japan28, the USA29, and Germany30.
Comparing the two options for ovarian tissue cryopreservation (OTC)-vitrification versus the standard procedure of slow freezing-results are partially conflicting in current meta-analyses16. Several factors may have contributed to this, as current vitrification protocols vary greatly. These differences include the choice of cryoprotectant or combination of protectants, their concentration, the composition of the OTC media, the size of tissue fragments, and the device used as a tissue carrier. Accordingly, there is no standardized warming protocol.
As the authors found a method that yields convincing results in terms of handling, viability, apoptosis onset, release of angiogenic factors, and even a report of a birth after reimplantation9,27, a very detailed description of the protocol is provided. The described method offers a valid and effective protocol that may contribute to the standardization of vitrification of ovarian tissue.

<meta charset="utf8"></head><body>著者スポットライト:卵巣組織ガラス化とIn vitro卵胞成長による若年女性がん患者の妊孕性保存の推進

ガラス質の凝集状態を達成するための卵巣皮質組織のガラス化

The scope of our research group is fertility protection in young female cancer patient. It is important to improve existing protocols to increase the chances of a later pregnancy. That includes the vitrification of ovarian tissue and also the in vitro growth of follicles With the in vitro growth of ovarian follicular.The crucial point is the multi-step tissue culture approach prior to the follicular isolation procedure as ovarian cortex tissue is very dense and limits follicular growth. Tissue vitrification is more complex than the slow freezing procedure, but it simplifies the whole process and has other advantages as well. We have shown that vitrification is a significant alternative to the common slow freezing protocol, which is supported by the previously reported five successful deliveries.The stress of freezing and thawing as well as the undersupply after reimplantation, together with the reperfusion damage impacts follicular survival and leads to a significant decrease in the number of the follicles. Limiting the success of the therapy option, vitrification is thought to preserve the follicles with a similar or even greater success compared to slow freezing approaches. We aim, therefore, to bring vitrification to a pre-clinical setting, that will provide all cryobank the possibility for tissue cryo-preservation, despite any personal and financial limitations.

Ovarian tissue cryopreservation (OTC) is an important option for fertility preservation. For patients whose gonadotoxic treatments cannot be postponed or ...

vitrification-ovarian-cortex-tissue-to-achieve-glassy-state

Research

JoVE Journal

Medicine

Vitrification of Ovarian Cortex Tissue to Achieve a Glassy State of Aggregation

817 ビュー.  University Hospital of Bonn. 当研究グループでは、若年女性がん患者の妊孕性保護に取り組んでいます。既存のプロトコルを改善して、後の妊娠の可能性を高めることが重要です。これには、卵巣組織のガラス化と、卵胞のin vitro成長と卵巣卵胞のin vitro成長が含まれます。重要な点は、卵巣皮質組織が非常に密集し、卵胞の成長を制限するため、卵胞隔離手順の前に?...