Cloning using donor nuclei from differentiated fetal and adult cells

Abstract

An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.

1 - 77 . (canceled)  
     
     
         78 . A method of producing a CICM cell line, comprising: 
 (i) inserting a desired mammalian somatic cell or mammalian somatic cell nucleus into an enucleated mammalian oocyte of the same species as the somatic cell or cell nucleus, under conditions suitable for the formation of a nuclear transfer (NT) unit;    (ii) activating the resultant nuclear transfer unit;    (iii) culturing said activated nuclear transfer unit until greater than the 2-cell developmental stage and no greater than the 400-cell developmental stage; and    (iv) culturing cells obtained from said cultured NT unit to obtain a CICM cell line which is cultured under conditions that maintain said CICM cell line in an undifferentiated state.    
     
     
         79 - 106 . (canceled)