|
|
Blood, Vol. 114, Issue 9, 1764-1767, August 27, 2009
Definitive proof for direct reprogramming of hematopoietic cells to pluripotency
Blood Okabe et al.
114: 1764
Supplemental materials for: Okabe et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 122 KB)
- Table S1. Primers used for PCR analyses (PDF, 41.1 KB)
- Figure S1. Generation of iPSCs derived from BM hematopoietic cells (JPG, 382 KB)
-
(A) Appearance of cells during the process of iPSC induction from BM HSPCs. Bar represents 100 µm. (B) SSEA-1 expression in BM hematopoietic cell-derived iPS cells (BM-derived iPSCs). Representative flow cytometric plots are shown for established BM-derived iPSCs (right) together with those for whole BM (left) and ES cells (center). Note the expression of SSEA-1 in BM-derived iPS cells at levels comparable to those in ES cells and the absence of SSEA-1 expression in whole BM cells.
- Figure S2. Assessment of iPSC factor genes in sHSC-iPSCs (JPG, 234 KB)
-
(A) PCR analysis shows the presence of all iPSC proviruses in each sHSC-iPS clone. H2O: No-template control; Control: Positive control using each vector DNA as template. (B) RT-PCR analysis using specific primer sets to examine expression of the 4 iPSC factors. In each sHSC-iPS clone, transcripts from the endogenous genes are detectable (Endo), while those from the retroviruses are not (Tg). RT (−): No–reverse-transcriptase control; Control: Jurkat cells transduced with each retrovirus vector. Note the near-absence of Tg amplicons in test samples after 35-cycle amplification.
- Figure S3. Assessment of expression of pluripotency markers in sHSC-iPSCs (JPG, 252 KB)
-
(A) RT-PCR analysis showing ES marker gene expression in sHSC-iPSC clones. H2O: No-template control; ES: ES cells as a positive control; RT (−): No–reverse-transcriptase control. (B) Nanog expression in sHSC-iPSC clones shown by immunostaining. NC: Negative control with isotype control antibodies. Nuclei were stained with DAPI. Bar represents 100 µm.
- Figure S4. Developmental potential of sHSC-iPSCs (JPG, 372 KB)
-
Shown are photomicrographs of histological sections of teratomas derived from sHSC-iPSC clone #9-5. Bar represents 100 µm.
- Figure S5. Assessment of iPSC factor genes in pHPC-iPSC (JPG, 252 KB)
-
(A) PCR analysis shows the presence of all iPSC factors in each pHPC-iPSC clone. H2O: No-template control; Control: Positive control using each vector DNA as template. (B) RT-PCR analysis using specific primer sets to examine expression of the 4 iPSC factors. In each pHPC-iPSC clone, transcripts from the endogenous genes are detectable (Endo), while those from the retroviruses are not (Tg). RT (−): No–reverse-transcriptase control; Control: Jurkat cells transduced with each retrovirus vector. Note the near-absence of Tg amplicons in test samples after 35-cycle amplification.
|
|
