Oil immersion technique for cellular microencapsulation

R. Nurhayati, W. Mubarok, Rafianto Dwi Cahyo, Kamila Alawiyah
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Abstract

Recent advances in tissue engineering application for medical intervention has created a need for three-dimensional (3D) cellular microenvironment in the form of cellular microencapsulation. Cellular microencapsulation can be used in several applications such as in vitro drug screening as well as cellular transplantation. It is important to note, however, that despite all the benefit from the cells, the capsule itself also has a major role. Significant impact and even clinical risks can present if the capsule is not biocompatible or even toxic. Hence, careful consideration on the materials and more importantly the fabrication process of the capsule is needed. In this work, we presented a method for cellular encapsulation using oil immersion technique. Briefly, suspension of K562 cells and collagen was injected into the mineral oil on a cell strainer. Microcapsule then washed with PBS and immersed with culture medium. Photomicrograph analysis was performed to evaluate the shape as well as to measure the volume of the microcapsule. Immunohistochemistry staining with Propidium Iodide (PI) was used to confirm K562 entrapment within the microcapsule. Cellular microencapsulation using oil immersion technique resulted in a rounded sphere. The increased volume of cells-hydrogel suspension resulted in bigger diameter and volume of the capsule. Due to the nature of the immersion process, a layer of oil could be found outside the capsule. The volume of the oil on the border of the capsule also increased with higher hydrogel-cells suspension. PI-stained K562 cells were found within the microcapsule, confirming that K562 cells were indeed entrapped within the microcapsule. In conclusion, oil immersion technique provides a quick, easy, and feasible method to produce cellular microencapsulation
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细胞微胶囊化的油浸技术
组织工程应用于医疗干预的最新进展创造了以细胞微胶囊形式的三维(3D)细胞微环境的需求。细胞微囊化可以用于多种应用,如体外药物筛选以及细胞移植。然而,值得注意的是,尽管这些细胞有很多好处,但胶囊本身也起着重要作用。如果胶囊不具有生物相容性甚至有毒,则会产生重大影响甚至临床风险。因此,需要仔细考虑材料,更重要的是胶囊的制造工艺。在本研究中,我们提出了一种利用油浸技术进行细胞包封的方法。简单地说,将K562细胞和胶原的悬浮液通过细胞过滤器注射到矿物油中。然后用PBS洗涤微胶囊,用培养基浸泡。显微分析进行了评估形状以及测量微胶囊的体积。免疫组织化学染色碘化丙啶(PI)证实K562包埋在微胶囊内。细胞微胶囊化采用油浸技术,形成一个圆形球体。细胞-水凝胶悬浮液体积增大,囊直径增大,囊体积增大。由于浸没过程的性质,可以在胶囊外面发现一层油。水凝胶细胞悬浮液浓度越高,胶囊边缘的油体积也越大。在微胶囊内发现pi染色的K562细胞,证实K562细胞确实包被在微胶囊内。总之,油浸技术是一种快速、简便、可行的制备细胞微胶囊的方法
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