医药/药品类部分翻译案例
狭窄或闭塞的动脉旁路移植术在当前的血管疾病中仍是一种重要的临床干预手段。此类手术通常使用患者自体静脉/动脉,或由涤纶或膨体聚四氟乙烯(ePTFE)等制作的人工血管实施 。
Stenosis or occlusion arterial bypass grafting is still an important clinical intervention means for current vascular diseases. Such surgeries are often made using the artificial blood vessel made of the patient’s autogenous vein or artery or dacron or ePTFE, etc.
使用患者自体血管进行移植往往受限于患者自身的病情或无法取得合适移植的血管组织 , 因此,在人工血管领域进行了长期广泛的努力。然而,人工血管普遍存在的未解决问题是:易形成血栓、发生再狭窄、顺应性失配、长期开放性低、长期抗凝治疗差以及因缺少内皮细胞而导致的生命周期有限 。至今没有任何材料能被证明适合生成直径小于6毫米的移植人造血管,且用于替代小直径动脉(如冠状动脉和膝下腹缘血管)时的开放率(12个月跟踪的平均值为51%)也不尽如人意。
Grafting using the patient’s autogenous vein or artery is often limited by the patient’s own condition or failure to obtain the vascular tissue that is suitable for grafting. Therefore, a long term and extensive efforts have been made in the field of artificial blood vessel. However,the prevailing unsolved problems of artificial blood vessel are as follows: easy formation of thrombus, generation of restenosis, compliance mismatch, long-term low openness, long-term weak anticoagulant therapy and limited life circle due to lack of endothelial cells. So far, no materials can be proved to be suitable for generate an artificial blood vessel for grafting with a diameter of less than 6mm, and the rate of openness (average tracking value of 12 months is 51% ) when it is used to replace the small diameter arteries (such as coronary artery and infragenicular wall blood vessel) is also not satisfactory.
我公司采用自体脂肪组织为来源的间充质干细胞在移植后的人工血管内部生成了内皮细胞这一方法,在解决以上挑战方面取得了进展。
Our company uses mesenchymal stem cell with autologous adipose tissue as the source to generate endothelial cells in the artificial blood vessel grafted, which makes a progress in solving the above challenges.
3.3 P1代猪来源脂肪间充质干细胞的培养方法
3.3 Culture Method for P1 Generation Pig Source Adipose Tissue-derived Stromal Cells
(1)原代培养脂肪间充质干细胞:使用酶消化法,通过I型胶原酶将脂肪间充质干细胞从脂肪组织中分离,随后通过过滤、离心的方法收集提取的脂肪间充质干细胞,接种于T75培养瓶中,使用含10%胎牛血清的DMEM/F12培养基,于37 ℃、5%CO2的条件下进行培养,获得P0代猪来源脂肪间充质干细胞(具体请详见附件一,1:《脂肪间充质干细胞培养用液配制操作规程》;2:《脂肪间充质干细胞P0分离培养标准操作规程》;3:《脂肪间充质干细胞培养常规操作标准操作规程》);
(1) Primary culture of adipose tissue-derived stromal cells : By enzyme digestion, separate the adipose tissue-derived stromal cells from the adipose tissue with I-type collagenase, and then collect and extract the adipose tissue-derived stromal cells by filtering and centrifugation, and inoculate them to T75 culture bottle, and culture them with DMEM/F12 culture medium containing 10% fetal calf serum at 37 ℃ and 5%CO2 to obtain P0 generation pig source adipose tissue-derived stromal cells (for details, see Appendix I, 1: Operation Procedures for Preparation of Adipose Tissue-derived Stromal Cells Culture Solution; 2:Standard Operation Procedures for P0 Isolated Culture of Adipose Tissue-derived Stromal Cells; 3 Standard Operation Procedures for Normal Operation of Adipose Tissue-derived Stromal Cells Culture);
(2)待培养的P0代猪来源脂肪间充质干细胞汇合度达到80%-90%时,去除细胞培养上清,使用CTSTM TrypLETM Select Enzyme消化细胞,离心、收集细胞,对细胞进行传代操作,获得P1代细胞(具体请详见附件一,4:《脂肪间充质干细胞传代标准操作规程》);
(2) When the confluence of P0 generation pig source adipose tissue-derived stromal cells to be cultured reaches 80%-90%, remove the cell cultural supernatant, digest, centrifuge and collect the cells with CTSTM TrypLETM Select Enzyme, and conduct a passage operation for the cells, to obtain P 1 generation cells (For details, see Appendix I, 4: Standard Operation Procedures for Passage of Adipose Tissue-derived Stromal Cells);
(3)P1代细胞接种于T-75培养瓶中,使用Lonza培养基,于37 ℃、5% CO2条件下对P1代细胞进行扩增培养,待细胞融合度达到90%时,开始细胞冻存及入库操作。在入库之前,细胞需进行质量检测(具体请详见附件一,3:《脂肪间充质干细胞培养常规操作标准操作规程》)。
(3) P1 generation cells are inoculated in T-75 culture bottle, and multiplication culture is made for P1 generation cells at 37 ℃ and 5% CO2, and cell cryopreservation and in-stockroom operation will be started when the confluence of cells reaches 90%. Before being put into storage, quality inspection shall be made for the cells (For details, see Appendix I, 3: Standard Operation Procedures for Normal Operation of Adipose Tissue-derived Stromal Cells Culture).
3.3.2 制备血管
3.3.2 Preparation of Blood Vessel
(1)启动3D生物打印机制备程序;
(1) Start the 3D bio-printer preparation procedure;
(2)运行喷头A,打印生物墨汁A,使其在旋生仪的旋转杆上形成长度为50-100 mm(长度根据实际需要而定)的血管结构体,厚度约1 mm;
(2) Operate nozzle A to print biological ink A and make it form a 50-100 mm (length depending on actual need) blood vessel structural body with a thickness of about 1mm on the rotating bar of rotating generation instrument;
(3)运行B喷头,在生物墨汁A形成的血管结构体上均匀喷涂生物墨汁B;
(3) Operate nozzle B to evenly spray biological ink B on the blood vessel structural body formed by geological ink A;
(4)将尺寸相匹配的人工血管套在血管结构体外表面,进行组装,通过生物墨汁B将两者粘合成一体,形成3D生物打印血管;
(4) Place the artificial blood vessel with matched dimensions on the outer surface of blood vessel structural body for assembly, and glue the two as a whole with biological ink B, to form a 3D biological printing blood vessel.
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