LaIuppa J A, McAdams T A, Papoutsakis E T, Miller W M
Department of Chemical Engineering, Northwestern University, Evanston, Illinois 60208-3120, USA.
J Biomed Mater Res. 1997 Sep 5;36(3):347-59. doi: 10.1002/(sici)1097-4636(19970905)36:3<347::aid-jbm10>3.0.co;2-b.
Ex vivo expansion of hematopoietic cells is important for applications such as cancer treatment, gene therapy, and transfusion medicine. While cell culture systems are widely used to evaluate the biocompatibility of materials for implantation, the ability of materials to support proliferation of primary human cells in cultures for reinfusion into patients has not been addressed. We screened a variety of commercially available polymer (15 types), metal (four types), and glass substrates for their ability to support expansion of hematopoietic cells when cultured under conditions that would be encountered in a clinical setting. Cultures of peripheral blood (PB) CD34+ cells and mononuclear cells (MNC) were evaluated for expansion of total cells and colony-forming unit-granulocyte monocyte (CFU-GM; progenitors committed to the granulocyte and/or monocyte lineage). Human hematopoietic cultures in serum-free medium were found to be extremely sensitive to the substrate material. The only materials tested that supported expansion at or near the levels of polystyrene were tissue culture polystyrene, Teflon perfluoroalkoxy, Teflon fluorinated ethylene propylene, cellulose acetate, titanium, new polycarbonate, and new polymethylpentene. MNC were less sensitive to the substrate materials than the primitive CD34+ progenitors, although similar trends were seen for expansion of the two cell populations on the substrates tested. CFU-GM expansion was more sensitive to substrate materials than was total cell expansion. The detrimental effects of a number of the materials on hematopoietic cultures appear to be caused by protein adsorption and/or leaching of toxins. Factors such as cleaning, sterilization, and reuse significantly affected the performance of some materials as culture substrates. We also used PB CD34+ cell cultures to examine the biocompatibility of gas-permeable cell culture and blood storage bags and several types of tubing commonly used with biomedical equipment. While many of the culture bag materials gave satisfactory results, all of the tubing materials severely inhibited total cell and CFU-GM expansion. Taken together, our results show that many materials approved for blood contact or considered biocompatible are not suitable for use with hematopoietic cells cultured in serum-free medium. As hematopoietic cultures are scaled up for a variety of clinical applications, it will be essential to carefully examine the biocompatibility of all materials involved.
造血细胞的体外扩增对于癌症治疗、基因治疗和输血医学等应用非常重要。虽然细胞培养系统被广泛用于评估植入材料的生物相容性，但材料在支持原代人类细胞增殖以供重新输注到患者体内的培养中的能力尚未得到研究。我们筛选了多种市售的聚合物（15种）、金属（4种）和玻璃基质，以评估它们在临床环境中遇到的条件下培养时支持造血细胞扩增的能力。对外周血（PB）CD34+细胞和单核细胞（MNC）培养物的总细胞扩增和集落形成单位 - 粒细胞单核细胞（CFU - GM；致力于粒细胞和/或单核细胞谱系的祖细胞）进行了评估。发现在无血清培养基中的人类造血培养物对基质材料极其敏感。测试的唯一能在聚苯乙烯水平或接近聚苯乙烯水平支持扩增的材料是组织培养聚苯乙烯、全氟烷氧基聚四氟乙烯、氟化乙烯丙烯聚四氟乙烯、醋酸纤维素、钛、新型聚碳酸酯和新型聚甲基戊烯。MNC对基质材料的敏感性低于原始CD34+祖细胞，尽管在测试的基质上两种细胞群体的扩增呈现相似趋势。CFU - GM扩增对基质材料的敏感性高于总细胞扩增。许多材料对造血培养的有害影响似乎是由蛋白质吸附和/或毒素浸出引起的。清洁、灭菌和再利用等因素显著影响了一些材料作为培养基质的性能。我们还使用PB CD34+细胞培养物来检查透气细胞培养和血液储存袋以及几种常用于生物医学设备的管材的生物相容性。虽然许多培养袋材料给出了令人满意的结果，但所有管材材料都严重抑制了总细胞和CFU - GM的扩增。综上所述，我们的结果表明，许多被批准用于血液接触或被认为具有生物相容性的材料并不适合用于无血清培养基中培养的造血细胞。随着为各种临床应用扩大造血培养规模，仔细检查所有相关材料的生物相容性将至关重要。
