Perbandingan Ko-kultur 2D dan 3D dengan Metode Hanging Drop untuk Menghasilkan Micro-environment yang Lebih Relevan Secara Klinis
Perbandingan Ko-kultur 2D dan 3D dengan Metode Hanging Drop untuk Menghasilkan Micro-environment yang Lebih Relevan Secara Klinis
复制标题
Perbandingan Ko-kultur 2D dan 3D dengan Metode Hanging Drop untuk Menhasilkan Microenvironment yang Lebih Relevan Secara Klinis
DOI：
10.23886/ejki.5.7701
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Iqbal Fasha
中科院分区：
文献类型：
--
作者：
Radiana Dhewayani Antarianto;Wahyunia Likhayati Septiana;A. Jusuf;Atikah Chalida Barasila;Jeannne Adiwinata Pawitan;Iqbal Fasha
2-二维共培养物与体内微环境的相似性有限，而三维共培养物形成的微团与体内微环境更相似，因此在生物医学研究中更有用。本研究旨在比较脐带血管来源的干细胞和肝星状细胞的2D共培养物与3D共培养物，采用悬滴法进行，以评估细胞形态和微团的球状体形成。本体外研究于2015年9月至2016年10月在人类病毒学和癌症病理学研究所（IHVCB）UI和组织学实验室进行，使用从人脐带血（UCB）和LX-2细胞系（人肝星状细胞）中分离的干细胞。用MACS CD 34分选人脐带血，并通过流式细胞术分析CD 34+细胞的百分比。采用悬滴法进行干细胞共培养（UCB）或脐带与L2的二维共培养和三维共培养。对每组共培养物进行一式三份实验。结果表明，与单培养相比，二维共培养和三维悬滴培养的形态有所不同。在2D共培养中，存在微团形成，而 在2D单一培养中，未形成微团。在3D悬滴中，与3D悬滴单一培养物相比，存在较小的球体。悬滴法二维和三维共培养细胞与单培养细胞相比，其形态发生了表型变化，细胞聚集在微团中。
2-dimensional (2D) co-cultures show limited resemblence with in vivo microenvironment while 3-dimensional (3D) co-cultures form a micromass which is more similar with in vivo microenvironmentthus it would be more useful in biomedical research. This study was intended to compare 2D co-cultures with 3D co-cultures of stem cells derived from umbilical cord blood vessels and hepatic stellate cells, conducted with hanging drop method to assess the cell morphology and the formation of spheroid from the micromass.. This in vitrostudy was conducted at Institute of Human Virology and Cancer Pathobiology (IHVCB) UI and histology laboratory from September 2015 to October 2016 using stem cells which were isolated from human umbilical cord blood (UCB) and LX-2 cell line (human hepatic stelate cells). Human umbilical cord blood was sorted with MACS CD34 and percentage of CD34+ cells were analyzed by flowcytometry. Stem cell co-cultures (UCB) or umbilical cord and L2 was did by hanging drop methods for 2D co-culture and 3D co-culture. Triplicates experiments were performed for each set of co-culture. The results showed the difference in the morphology of 2D co-culture and 3D hanging drop compared to monoculture. In the 2D co-culture there was a micromass formation, whereas in the 2D monoculture, the micromass was not formed. In the 3D hanging drop there was a smaller spheroid compared to the 3D hanging drop monoculture. The morphology of 2D and 3D co-culture cells with hanging drop method in comparison with monoculture cells showed phenotypic changes of cells which were incorporated together in the micromass.