大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

目的 探讨大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植治疗肾上腺功能不全的可行性.方法 分离SD大鼠(出生3 d内的乳鼠)的肾上腺细胞,置于自制鼠尾Ⅰ型胶原凝胶中培养,1周后移植到双侧肾上腺切除的成年雄性SD大鼠肾被膜下.术后每周取血1次,测量血浆皮质酮和醛固酮的水平;移植8周后处死大鼠,观察移植物的组织学特点;肾上腺细胞的增殖情况及CYP11B1和CYP11B2蛋白的表达情况;并与单纯肾上腺细胞移植组和正常对照组相比较.结果 大鼠移植经胶原凝胶三维培养的肾上腺细胞后,80%(12/15)存活达8周;手术后血浆皮质酮水平显著高于同期单纯肾上腺细胞移植组,移植6～8周时,与正常对照组已无显著性差异;其醛固酮水平术后恢复缓慢,与单纯肾上腺细胞移植组的差异无统计学意义;移植后肾上腺细胞在胶原内生长良好,增殖率高,95%为束状带细胞,表达CYP11B1,而表达CYP11B2的球状带细胞仅占5%,未见炎症细胞浸润.结论 肾上腺细胞经胶原凝胶三维培养后移植能形成有功能的血管化肾上腺组织,长期效果理想,有望成为临床治疗肾上腺功能不全的有效方法.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.     

大鼠肾上腺细胞经胶原凝胶三维培养后进行同种移植的实验研究

Objective To investigate a novel method of adrenocotical cells transplantation. Methods Adrenal glands of neonate rats were dissociated into adrenal cortical cells. Cells were cultured in self-made collagen type Ⅰ gel for one week, and then transplanted under the renal capsule of bilateral adrenalectomy mate rats. Blood samples were collected per week after surgery. Animals were sacrificed at the 8th week, and histological characteristics of the allografts, proliferation of transplanted cells, CYP11B1 and CYP11B2 expression as well as plasma aldosterone and corticosterone were observed. Results There were totally 15 rats receiving collagen gel transplantation, and 12 survived 8 weeks post-operation. The plasma cortocosterone level in collagen gel transplantation group was significantly higher than in adrenocortical cell transplantation group, and reached the normal level from 6th to 8th week, but the change in plasma aldosterone level in collagen gel transplantation group was similar to that of adrenocortical cell transplantation group. The adrenocortical cells cultured in gel grew well, and had a high proliferation rate. 95% of them were fasciculata cells which expressed CYP11B1, and the rest were glomerulosa cells which expressed CYP11B2. No inflammatory cell infiltration was observed. Conclusion The proliferation and function of adrenocortical cells could be promoted when they were cultured in collagen gel.