Patrick C W, Chauvin P B, Hobley J, Reece G P
Laboratory of Reparative Biology and Bioengineering, Department of Plastic Surgery, University of Texas M.D. Anderson Cancer Center, Houston, Texas 77030, USA.
Tissue Eng. 1999 Apr;5(2):139-51. doi: 10.1089/ten.1999.5.139.
Adipose tissue equivalents have not been addressed as yet despite the clinical need in congenital deformities, posttraumatic repair, cancer rehabilitation, and other soft tissue defects. Preadipocytes were successfully harvested from rat epididymal fat pads of Sprague-Dawley and Lewis rats and expanded ex vivo. In vitro cultures demonstrated full differentiation of preadipocytes into mature adipocytes with normal lipogenic activity. The onset of differentiation was well-controlled by regulating preadipocyte confluency. Poly(lactic-co-glycolic) acid (PLGA) polymer disks with 90% porosity, 2.5 mm thick, 12 mm diameter, pore size range of 135-633 microm were fabricated and seeded with preadipocytes at 10(5) cells/mL. Disks in vitro demonstrated fully differentiated mature adipocytes within the pores of the disks. Short-term in vivo experiments were conducted by implanting preseeded disks subcutaneously on the flanks of rats for 2 and 5 weeks. Histologic staining of harvested disks with osmium tetroxide (OsO4) revealed the formation of adipose tissue throughout the disks. Fluorescence labeling of preadipocytes confirmed that formed adipose tissue originated from seeded preadipocytes rather than from possible infiltrating perivascular tissue. This study demonstrates the potential of using primary preadipocytes as a cell source in cell-seeded polymer scaffolds for tissue engineering applications.
尽管先天性畸形、创伤后修复、癌症康复及其他软组织缺损存在临床需求，但脂肪组织替代物尚未得到研究。已成功从Sprague-Dawley大鼠和Lewis大鼠的附睾脂肪垫中获取前脂肪细胞，并在体外进行扩增。体外培养显示前脂肪细胞可完全分化为具有正常脂肪生成活性的成熟脂肪细胞。通过调节前脂肪细胞的汇合度可很好地控制分化的起始。制备了孔隙率为90%、厚度为2.5 mm、直径为12 mm、孔径范围为135 - 633微米的聚乳酸-乙醇酸共聚物（PLGA）聚合物圆盘，并以10(5)个细胞/毫升的密度接种前脂肪细胞。圆盘在体外显示其孔隙内有完全分化的成熟脂肪细胞。通过将预先接种细胞的圆盘皮下植入大鼠侧腹进行2周和5周的短期体内实验。用四氧化锇（OsO4）对收获的圆盘进行组织学染色，显示整个圆盘内形成了脂肪组织。前脂肪细胞的荧光标记证实，形成的脂肪组织源自接种的前脂肪细胞，而非可能浸润的血管周围组织。本研究证明了将原代前脂肪细胞用作细胞接种聚合物支架的细胞来源用于组织工程应用的潜力。
