SD大鼠毛乳头细胞体外培养及组织工程皮肤的构建

SD Rat Dermal Papilla Cell Culture In Vitro and Construction of A Tissue Engineered Skin

目的了解SD大鼠毛乳头细胞(DPCs)体外培养的生长特性,并用其构建组织工程双层皮肤。方法分离大鼠触须部获取完整毛囊,胶原酶消化获取毛乳头组织,组织块法培养毛乳头细胞并传代。以Ⅰ型胶原、α-平滑肌肌动蛋白(smooth muscle actin-α,SMA-α)、CK抗体免疫组化法鉴定细胞,MTT法检测并绘制细胞生长曲线。将第二代DPCs作为真皮种子细胞,角质形成细胞作为表皮种子细胞,用气-液面培养制备组织工程双层皮肤,HE染色观察其组织学结构。结果成功培养DPCs并传代,Ⅰ型胶原、SMA-α染色阳性;成功构建出含表皮与真皮层的组织工程双层皮肤。结论体外培养得到的DPCs与表皮结合较好,可作为组织工程皮肤的种子细胞。

Objective To study the growth ability and characteristics in vitro of SD rat dermal papilla cells, and use them to construct tissue engineered skin. Methods The follicles were isolated from rat whiskers and were digested by collagenase to obtain dermal papillas, dermal papilla cells were cultured by the method of tissue culture and were passaged. The biological features of cultured cells were investigated by immunohistochemical method to detect the expression of α - smooth muscle actin, collagen I and CK, and the cell cycle was detected by MTT. The second passage DPCs were seeded in collagen and the keratinocytes were seeded on the surface to constructed tissue full - thickness engineered skin by the air - liquid interface method, stained by HE. Results DPCs were cultivated and passaged successfully, collagen I and SMA - α were expressed; The full - thickness tissue engineered skin was constructed successfully. Conclusion The DPCs cultivated in vitro were beneficial for connecting with epidermis, and DPCs could act as seed cells of tissue engineered skin.