MSCs与CD34^＋单核细胞共培养联合脱细胞支架植入体内构建兔阴茎海绵体

Construction of rabbit tissue-engineered corpus cavernosum with co-culture of BM-MSCs and CD34^+MNCs and transplantation in vivo

目的研究将异体骨髓间充质干细胞(MSCs)和CD34^+单核细胞(MNCs)共培养体系种植于脱细胞海绵体支架上在兔体内构建阴茎海绵体组织的可行性。方法分别从兔骨髓与兔外周血中提取并鉴定MSCs与CD34^+MNCs,将两种细胞组分以107/mL的密度种植于制备好的兔阴茎海绵体脱细胞胶原支架上,同时将相同密度的MSCs、CD34^+MNCs分别种植于支架上,培养4天后植入一种兔阴茎海绵体缺损模型中。3个月后对工程化组织进行取材,行H&E及Masson三色染色等组织学检测。结果 MSCs和CD34^+MNCs共培养组近白膜处的海绵体保留了正常海绵体结构,而MSCs组、CD34+单核细胞组及空白支架组以疤痕增生为主,尽管较之CD34^+单核细胞组及空白支架组,间充质干细胞组疤痕组织中的血管数目较多(P<0.05)。结论我们的研究证明了将MSCs和CD34^+MNCs的共培养体系种植于脱细胞支架上植入兔体内可以构建出一段组织学上与原生海绵体相似的组织;CD34^+MNCs在体内主要通过协同MSCs而非单独发挥其促血管化的生物学效应。

Objective To investigate the feasibility of co-cultured mesenchymal stem cells （MSCs） and CD34＋mononuclear cells seeded in acellular matrix to construct a section of rabbit tissue-engineered eavernosum in vivo. Methods MSCs and CD34＋mononuclear cells （MNCs） were extracted from bone marrow and peripheral blood. After being identified, the two components of the cells were seeded in prepared aeellular corporal matrix alongside with MSCs and CD34 ＋ MNCs seeded in the same density respectively. The cell-laden matrix was transplanted into a rabbit model of penile interruption after culture for 4 days in vitro. Three months after implantation, animals were killed and the engineered tissue was collected for histological examination （H＆E staining and Masson＇s trichrome staining）. Results The engineered corporal tissue near tunica albuginea in MSCs and CD34＋MNCs cocultured group preserved the histological feather of intact corporal tissue. The other 3 groups, including MSCs group, CD34MNCs group and acellular matrix group, failed to preserve the corporal structure. Instead, they underwent fibrosis and formed sear tissue though samples from MSCs group showed more small vessels than from CD34＋ MNCs group and acellular matrix group （P〈0.05）. Conclusion Our work demonstrates the feasibility of co-implantation of MSCs and CD34＋ MNCs in acellular matrix to construct corporal tissue resembling to native one. CD34＋MNCs promote vascularization in vivo along with MSCs rather than alone.