Granicka L H, Wdowiak M, Kosek A, Swiezewski S, Wasilewska D, Jankowska E, Weryński A, Kawiak J
PAS, Institute of Biocybernetics and Biomedical Engineering, Warsaw, Poland.
Cell Transplant. 2005;14(5):323-30. doi: 10.3727/000000005783983043.
The purpose of the observations was the viability and quality evaluation of E. coli bacteria encapsulated in hollow fiber membranes (HF) in short in vivo and in vitro experiments. A polypropylene, surface-modified hollow fiber was applied for immunoisolation of E. coli bacteria transfected with a green fluorescent protein (E. coli GFPI). The presence of GFP fluorescence of organisms was assessed with the use of flow cytometry. The E. coli GFPIs were then observed for the period of 5 days in in vitro experiments in the culture medium. A single IPTG (isopropyl beta-D-1-thiogalactopyranoside) induction of GFP gene appeared to be adequate for an expression of GFP protein for 5 days. The GFP expression values observed for E. coli GFPs encapsulated in HF during culture in different culture media were comparable. The survival of E. coli GFPIs encapsulated in HF after 1, 2, 4, or 5 days of subcutaneous implantation into mice was evaluated. The explanted E. coli GFPIs exhibited mean expression 603 +/- 17 (n = 32) units of fluorescence during the implantation period. The values obtained were comparable for selected days of observation. It was observed that the membranes applied ensured the bacteria growth within the HF's space only.
这些观察的目的是在短期体内和体外实验中评估包裹在中空纤维膜（HF）中的大肠杆菌的活力和质量。一种表面改性的聚丙烯中空纤维用于对转染了绿色荧光蛋白的大肠杆菌（大肠杆菌GFPI）进行免疫隔离。使用流式细胞术评估生物体中GFP荧光的存在。然后在体外实验中，将大肠杆菌GFPI在培养基中培养5天进行观察。GFP基因的单次IPTG（异丙基-β-D-1-硫代半乳糖吡喃糖苷）诱导似乎足以使GFP蛋白表达5天。在不同培养基中培养期间，观察到包裹在HF中的大肠杆菌GFP的GFP表达值具有可比性。评估了将包裹在HF中的大肠杆菌GFPI皮下植入小鼠1、2、4或5天后的存活率。在植入期间，取出的大肠杆菌GFPI表现出平均荧光表达量为603±17（n = 32）个荧光单位。在选定的观察日获得的值具有可比性。观察到所应用的膜仅确保细菌在HF空间内生长。
