The use of human serum in supporting the in vitro and in vivo proliferation of human conjunctival epithelial cells
The use of human serum in supporting the in vitro and in vivo proliferation of human conjunctival epithelial cells
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DOI：
10.1136/bjo.2004.055046
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发表时间：
2005-06-01
影响因子：
4.1
通讯作者：
Beuerman, RW
中科院分区：
文献类型：
--
作者：
Ang, LPK;Tan, DTH;Beuerman, RW
目的：目的：探讨人血清（HS）对人结膜上皮细胞体外和体内增殖的促进作用，并与胎牛血清（FBS）和牛垂体提取物（BPE）进行比较。殖民地形成率（CFE）、溴脱氧尿苷（BrdU）ELISA增殖试验和细胞代次。通过免疫染色和RT-PCR评价细胞的角蛋白（K4、K19和K3）和MUC 5AC表达。将羊膜上构建的结膜移植到严重联合免疫缺陷（SCID）小鼠体内10天，并进行组织学分析。（CFE，6.7%（SD 1.8%）; BrdU吸光度，0.86（0.16））与补充FBS的（CFE，9.3%（1.8%）; BrdU吸光度，1.11（0.18））和BPE补充的培养物（CFE，5.9（1.5）; BrdU吸光度，0.65（0.12））。补充HS、FBS和BPE的培养基的杯状细胞密度分别为52个细胞/cm（2）、60个细胞/cm（2）和50个细胞/cm（2）。HS补充文化形成分层上皮片在体内transplantation.Conclusions：培养在HS补充文化的结膜上皮细胞的增殖能力是FBS和BPE补充文化相媲美。当培养用于临床移植的细胞时，从培养系统中去除动物材料是有利的。
Aim: To evaluate the use of human serum (HS) in supporting the in vitro and in vivo proliferation of human conjunctival epithelial cells, and compare it with fetal bovine serum (FBS) and bovine pituitary extract (BPE).Methods: Conjunctival epithelial cells were cultivated in media supplemented with HS (5%, 10%), FBS (5%, 10%), and BPE (70 mg/ml, 140 mg/ml). The colony forming efficiency (CFE), bromodeoxyuridine (BrdU) ELISA proliferation assay, and cell generations were analysed. Cells were evaluated for keratin (K4, K19, and K3) and MUC5AC expression by immunostaining and RT-PCR. Conjunctival equivalents constructed on amniotic membranes were transplanted onto severe combined immune deficient (SCID) mice for 10 days and analysed histologically.Results: The proliferation assays of HS supplemented cultures (CFE, 6.7% (SD 1.8%); BrdU absorbance, 0.86 (0.16)) were comparable to FBS supplemented (CFE, 9.3% (1.8%); BrdU absorbance, 1.11 (0.18)) and BPE supplemented cultures (CFE, 5.9 (1.5); BrdU absorbance, 0.65 (0.12)). Goblet cell densities for HS, FBS, and BPE supplemented media were 52 cells/cm(2), 60 cells/cm(2), and 50 cells/cm(2), respectively. HS supplemented cultures formed stratified epithelial sheets in vivo following transplantation.Conclusions: The proliferative capacity of conjunctival epithelial cells cultivated in HS supplemented cultures was comparable to FBS and BPE supplemented cultures. The elimination of animal material from the culture system is advantageous when cultivating cells for clinical transplantation.