Isachenko V, Isachenko E, Mallmann P, Rahimi G
Department of Obstetrics and Gynecology, Cologne University, Cologne, Germany.
Clin Lab. 2012;58(11-12):1293-300.
The positive effect of cooling on tissue cells is known. The aim of this research was to study the intensiveness of neo-vascularisation and follicular development in ovarian tissue after 24 hours cooling to 5 degrees C before cryopreservation.
Fifty six pieces from 7 patients were divided into the following four groups: Group 1: pieces cultured just after operation, Group 2: pieces cooled after operation to 5 degrees C for 24 hours and then cultured, Group 3: pieces frozen-thawed just after operation and then cultured, Group 4: pieces cooled after operation to 5 degrees C for 24 hours, frozen, thawed, and then cultured. Culture of ovarian pieces was performed in a chorioallantoic membrane (CAM)-system for 5 days. The efficacy of the tissue cooling was evaluated by the development of follicles and intensiveness of neo-vascularisation (by Desmin).
For Group 1, 2, 3, and 4, mean density of follicles per 1 mm3 was 10.1, 11.1, 9.8, and 12.0, respectively (P1-2, 3-4 < 0.05). For these groups 91%, 92%, 90%, and 90% preantral follicles were morphologically normal (P1-2, 3-4 > 0.1). The immunohistochemical analysis showed that the intensiveness of neo-vascularisation observed in ovarian tissue of Group 2 (pre-cooled before culture) and Group 4 (pre-cooled before cryopreservation) was drastically increased.
The 24 hour cooling to 5 degrees C before cryopreservation is beneficial for cryopreservation of human ovarian tissue.
冷却对组织细胞的积极作用是已知的。本研究的目的是探讨在冷冻保存前将卵巢组织冷却至5℃ 24小时后新血管形成和卵泡发育的强度。
将7例患者的56块组织分为以下四组：第1组：术后立即培养的组织块；第2组：术后冷却至5℃ 24小时后再培养的组织块；第3组：术后立即冷冻解冻后再培养的组织块；第4组：术后冷却至5℃ 24小时，冷冻，解冻，然后培养。卵巢组织块在绒毛尿囊膜（CAM）系统中培养5天。通过卵泡发育和新血管形成强度（通过结蛋白）评估组织冷却的效果。
第1、2、3和4组每1立方毫米卵泡的平均密度分别为10.1、11.1、9.8和12.0（P1-2、3-4 < 0.05）。这些组中91%、92%、90%和90%的窦前卵泡形态正常（P1-2、3-4 > 0.1）。免疫组织化学分析表明，第2组（培养前预冷）和第4组（冷冻保存前预冷）卵巢组织中观察到的新血管形成强度显著增加。
冷冻保存前将卵巢组织冷却至5℃ 24小时有利于人类卵巢组织的冷冻保存。
