Lakshmanan Yegappan, Frimberger Dominic, Gearhart John D, Gearhart John P
Division of Pediatric Urology, Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
Urology. 2005 Apr;65(4):821-6. doi: 10.1016/j.urology.2004.11.022.
Human pluripotent stem cells have excellent regenerative abilities, making them attractive in bladder regeneration. Porcine small intestinal submucosa (SIS) is an established matrix well-suited to the urinary tract. We evaluated the ability of human embryonic germ (hEG) cell-derived stem cells to form a composite graft on SIS, grown alone or in combination with human bladder smooth muscle cells (SMCs) or urothelial (URO) cells.
The hEG cell-derived cell line SDEC had the best-fit profile for smooth muscle, as determined by its gene and protein expression. SDEC cells were seeded on SIS, either on its serosal or mucosal surface, and cultured for 7 or 14 days. Co-cultures of stem cells with URO cells or SMCs were also assessed under similar conditions. The grafts were analyzed by histologic examination for cell growth, morphology, and matrix penetration.
SDEC cells grew in a monolayer on SIS, with a marked increase in three-dimensional growth when co-cultured with URO cells or SMCs. Penetration of the matrix was evident when seeded on the rough surface of the SIS, progressing with time. Stratification of the cell layers occurred on the smooth side of the SIS.
This is the first description of hEG cell-derived stem cells in co-culture with bladder cells. hEG cell-derived stem cells grow well on SIS even when seeded at low concentrations in the presence of bladder cells (URO cells or SMCs). These composite grafts will be ideal to evaluate the in vivo functional characteristics of stem cells using an animal model of bladder regeneration.
人类多能干细胞具有出色的再生能力，使其在膀胱再生方面具有吸引力。猪小肠黏膜下层（SIS）是一种适用于泌尿道的成熟基质。我们评估了人类胚胎生殖（hEG）细胞来源的干细胞在SIS上单独生长或与人类膀胱平滑肌细胞（SMC）或尿路上皮（URO）细胞联合生长时形成复合移植物的能力。
通过基因和蛋白质表达确定，hEG细胞来源的细胞系SDEC最适合平滑肌细胞特征。将SDEC细胞接种在SIS的浆膜或黏膜表面，培养7天或14天。在相似条件下也评估了干细胞与URO细胞或SMC的共培养情况。通过组织学检查分析移植物的细胞生长、形态和基质穿透情况。
SDEC细胞在SIS上单层生长，与URO细胞或SMC共培养时三维生长显著增加。接种在SIS粗糙表面时，基质穿透明显，并随时间推移而进展。在SIS光滑面出现细胞层分层。
这是首次描述hEG细胞来源的干细胞与膀胱细胞共培养的情况。hEG细胞来源的干细胞即使在低浓度接种且存在膀胱细胞（URO细胞或SMC）的情况下，也能在SIS上良好生长。这些复合移植物将是使用膀胱再生动物模型评估干细胞体内功能特性的理想材料。
