INDUCED PLURIPOTENT STEM CELLS FROM HUMAN UMBILICAL CORD TISSUE-DERIVED CELLS
We have disclosed an induced pluripotent stem cell and the method of preparing the induced pluripotent stem cell from a human umbilical cord tissue-derived cell. More particularly, we have disclosed a human umbilical cord tissue-derived iPS cell which may be differentiated into cells of ectoderm, mesoderm, and endoderm lineages.
1 . An induced pluripotent stem cell comprising a reprogrammed human umbilical cord tissue-derived cell wherein the human umbilical cord tissue-derived cell is an isolated umbilical cord tissue cell isolated from human umbilical cord tissue substantially free of blood that is capable of self-renewal and expansion in culture, has the potential to differentiate into cells of other phenotypes, can undergo at least 40 doublings in culture, maintains a normal karyotype upon passaging, and has the following characteristics: expresses each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2, and HLA-A,B,C; does not express any of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, or HLA-DR,DP,DQ; and increased expression of a gene for each of interleukin 8; reticulon 1; and chemokine (C-X-C motif) ligand 3 relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an iliac crest bone marrow cell.
2 . The induced pluripotent stem cell of claim 1 , wherein the wherein the human umbilical cord tissue-derived cell further has the following characteristics: secretes each of the factors MCP-1, MIP1beta, IL-6, IL-8, GCP-2, HGF, KGF, FGF, HB-EGF, BDNF, TPO, RANTES and TIMP1; and
does not secrete any of the factors SDF-1alpha, TGF-beta2, ANG2, PDGFbb, MIP1a and VEGF.
3 . The induced pluripotent stem cell of claim 1 , wherein the induced pluripotent stem cell expresses TRA1-60, TRA1-81, SSEA3, SSEA4, and NANOG.
4 . The induced pluripotent stem cell of claim 1 , wherein the induced pluripotent stem cell is positive for alkaline phosphatase staining.
5 . The induced pluripotent stem cell of claim 1 , wherein the induced pluripotent stem cell differentiates into cells of ectoderm, mesoderm, and endoderm lineages.
6 . An induced pluripotent stem cell prepared by a method comprising the steps of:
providing a human umbilical cord tissue-derived cell, wherein the human umbilical cord tissue-derived cell is an isolated umbilical cord tissue cell isolated from human umbilical cord tissue substantially free of blood that is capable of self-renewal and expansion in culture, has the potential to differentiate into cells of other phenotypes, can undergo at least 40 doublings in culture, maintains a normal karyotype upon passaging, and has the following characteristics: expresses each of CD10, CD13, CD44, CD73, CD90, PDGFr-alpha, PD-L2, and HLA-A,B,C; does not express any of CD31, CD34, CD45, CD80, CD86, CD117, CD141, CD178, B7-H2, HLA-G, or HLA-DR,DP,DQ; and increased expression of a gene for each of interleukin 8; reticulon 1; and chemokine (C-X-C motif) ligand 3 relative to that of a human cell which is a fibroblast, a mesenchymal stem cell, or an iliac crest bone marrow cell;
transfecting the human umbilical cord tissue derived-cell with murine retroviruses, individually carrying constitutively expressed human transcription factors OCT4, SOX2, KLF4, and c-MYC,
culturing the transfected human umbilical cord tissue-derived cell, identifying an induced pluripotent stem cell,
isolating the human umbilical cord tissue-derived IPS cell, subculturing the induced pluripotent stem cell, and
providing a induced pluripotent stem cell.
7 . The method of claim 5 , wherein the murine retrovirus further carries p53-shRNA.