RECONSTITUTION OF CELLS BY FUSION OF CELL FRAGMENTS .1. MYOGENIC EXPRESSION AFTER FUSION OF MINICELLS FROM RAT MYOBLASTS (L6) WITH MOUSE FIBROBLAST (A9) CYTOPLASM
RECONSTITUTION OF CELLS BY FUSION OF CELL FRAGMENTS .1. MYOGENIC EXPRESSION AFTER FUSION OF MINICELLS FROM RAT MYOBLASTS (L6) WITH MOUSE FIBROBLAST (A9) CYTOPLASM
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DOI：
10.1016/0014-4827(78)90363-4
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发表时间：
1978-01-01
影响因子：
3.7
通讯作者：
COLEMAN, JR
中科院分区：
文献类型：
--
作者：
RINGERTZ, NR;KRONDAHL, U;COLEMAN, JR
在细胞松弛素b的作用下，将大鼠L6成肌细胞和小鼠A9成纤维细胞（HGPRT-）单层离心去核，用仙台病毒介导的去核大鼠成肌细胞的细胞核（小细胞）与去核小鼠成纤维细胞的细胞质融合，重建完整的细胞。根据细胞核和细胞质标记，将重组细胞增殖产生的菌落与完整亲本细胞类型区分开来。在5次重复实验中，融合后发现的所有菌落中约有70%来自重组细胞，30%来自污染小细胞制剂的完整大鼠成肌细胞，2个菌落被鉴定为亲本细胞类型之间的杂交。由重组细胞衍生的克隆形成肌管，产生肌球蛋白并形成骨骼肌典型的交叉条纹。因此，大鼠成肌细胞的成肌程序可以通过去核和重建过程持续存在，并且不会因暴露于小鼠成纤维细胞细胞质一段时间而明显改变。
Rat L6 myoblasts and mouse A9 fibroblasts (HGPRT-) were enucleated by centrifugation of monolayers in the presence of cytochalasin B. Intact cells were reconstituted by Sendai virus mediated fusion of nuclei (minicells) from enucleated rat myoblasts and cytoplasms from enucleated mouse fibroblasts. Colonies arising from proliferating reconstituted cells were distinguished from intact parental cell types on the basis of nuclear and cytoplasmic markers. In 5 replicate experiments, approximately 70% of all colonies found after fusion were derived from reconstituted cells, 30% arose from intact rat myoblasts contaminating the minicell preparations, and 2 colonies were identified as hybrids between the parental cell types. Clones derived from reconstituted cells formed myotubes which produced myosin and developed the cross-striated pattern typical of skeletal muscle. The myogenic program of the rat myoblast thus can persist through enucleation and reconstitution procedures, and is not obviously altered by a period of exposure to mouse fibroblast cytoplasm.