Lee S, Kim B Y, Yeo J E, Nemeno J G, Jo Y H, Yang W, Nam B M, Namoto S, Tanaka S, Sato M, Lee K M, Hwang H S, Lee J I
Regenerative Medicine Laboratory, Center for Stem Cell Research, Department of Biomedical Science and Technology, Institute of Biomedical Science & Technology (IBST), Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, Republic of Korea.
Transplant Proc. 2013 Oct;45(8):3108-12. doi: 10.1016/j.transproceed.2013.08.013.
Before cell or tissue transplantation, cells or tissues have to be maintained for a certain period in vitro using culture medium and methods. Most culture media contain substances such as pH indicators and buffers. It is not known whether some of these substances are safe for subsequent application in the transplantation of cells or tissues into the human body. We investigated culture media and methods with respect to the safety of the components in future transplantation applications.
A modified culture medium--medical fluid-based culture medium (FCM)--was designed by using various fluids and injectable drugs that are already currently permitted for use in clinical medicine. Medium components necessary for optimal cell growth were obtained from approved drugs. FCM was manufactured with adjusted final concentrations of the medium components similar to those in commercial Dulbecco's modified Eagle's medium (DMEM). In particular, 1029.40 mg/L amino acids, approximately 88.85 mg/L vitamins, 13,525.77 mg/L inorganic salts, and 4500 mg/L D-glucose comprise the high-glucose FCM. Next, human fat synovium-derived mesenchymal stem cells and rat H9c2 (2-1) cells were cultured under 2 conditions: (1) DMEM-high glucose (HG), an original commercial medium, and (2) optimized FCM-HG. We assessed the morphologies and proliferation rates of these cells.
We observed that FCM-HG was able to induce the growth of FS-MSC and commercially available H9c2 cell. The morphologies and proliferation patterns of these cells cultured under FCM-HG showed no differences compared with cells grown in DMEM-HG.
Our data suggest that FCM, which we developed for the first time according to the concept of drug repositioning, was a useful culture medium, especially in cultured cells intended for human cell transplantation.
在细胞或组织移植之前，必须使用培养基和培养方法在体外将细胞或组织维持一定时间。大多数培养基都含有诸如pH指示剂和缓冲剂之类的物质。尚不清楚这些物质中的某些对于随后将细胞或组织移植到人体中是否安全。我们研究了培养基和培养方法在未来移植应用中成分的安全性。
通过使用目前已被批准用于临床医学的各种液体和注射用药物，设计了一种改良的培养基——医用液体基培养基（FCM）。最佳细胞生长所需的培养基成分取自已批准的药物。FCM的制造是将培养基成分的最终浓度调整至与市售杜氏改良 Eagle 培养基（DMEM）相似。特别是，高糖FCM包含1029.40mg/L氨基酸、约88.85mg/L维生素、13525.77mg/L无机盐和4500mg/L D-葡萄糖。接下来，将人脂肪滑膜来源的间充质干细胞和大鼠H9c2（2-1）细胞在两种条件下培养：（1）原始市售培养基DMEM-高糖（HG），以及（2）优化后的FCM-高糖（HG）。我们评估了这些细胞的形态和增殖率。
我们观察到FCM-高糖能够诱导脂肪滑膜间充质干细胞（FS-MSC）和市售H9c2细胞的生长。与在DMEM-高糖中生长的细胞相比，在FCM-高糖中培养的这些细胞的形态和增殖模式没有差异。
我们的数据表明，我们首次根据药物重新定位的概念开发的FCM是一种有用的培养基，特别是在用于人类细胞移植的培养细胞中。
