Li X, Abdi K, Mentzer S J
Division of Thoracic Surgery, Brigham and Women's Hospital, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115.
Hybridoma. 1992 Oct;11(5):645-52. doi: 10.1089/hyb.1992.11.645.
Alginate is a transparent polymer of guluronic and mannuronic acids that provide a favorable microenvironment for cell growth. Alginate gelation is calcium dependent and temperature independent. To facilitate the isolation of stable and productive antibody-producing hybridomas, we have developed a technique of cloning hybridomas in the three-dimensional alginate matrix. To provide cavities for hybridoma growth, we encorporated 10-15% (v/v) gelatin into the alginate prior to gelation. We have cloned more than 90 monoclonal antibody-producing hybridomas using the alginate matrix. The alginate matrix is readily reversible with the addition of a calcium chelator. The alginate matrix permits efficient cloning in limited incubator space, without the use of a feeder layer, and with minimal amount of medium. The transparent matrix also permits easy screening for clonality and growth.
藻酸盐是古洛糖醛酸和甘露糖醛酸的透明聚合物，可为细胞生长提供有利的微环境。藻酸盐凝胶化依赖于钙，与温度无关。为便于分离稳定且高产抗体的杂交瘤，我们开发了一种在三维藻酸盐基质中克隆杂交瘤的技术。为给杂交瘤生长提供腔室，我们在凝胶化前将10 - 15%（v/v）的明胶掺入藻酸盐中。我们已使用藻酸盐基质克隆了90多个产生单克隆抗体的杂交瘤。添加钙螯合剂后，藻酸盐基质易于逆转。藻酸盐基质允许在有限的培养箱空间内进行高效克隆，无需使用饲养层，且培养基用量最少。透明基质还便于克隆性和生长的筛选。
