Stem Cells: Minireview Presentation

Stem Cells Dr. Tai-ping Sun Biology 280S: Biotechnology & Genetic Engineering Jon Martin Joshua Mendoza-Elias Fall 2008

Outline: ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Properties of Stem Cells ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Where do SC come from? Embryonic Fetal Adult Berashis Stem Cells

For the love of cloning ,[object Object],[object Object],[object Object],[object Object],J.B. Gurdon (1962): S. African clawed frog ( Xenopus laevis ) Evidence from EvoDevoBio: The development of cloning techniques revealed organisms could be grown from one cell.

One of these things is not like the other. ,[object Object],[object Object],[object Object],[object Object],Proof of Concept: Clinical Procedure:

Mechanism of progenitor cells in the first steps of B-cell maturation. Receptors and ligands look familiar? ( c-kit , SCF )

Techniques for studying ES cells The more things change, the more the stay the same. ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

II. Techniques in Stem Cell Therapy

Therapeutic Cloning ,[object Object]

Problems with Therapeutic Cloning ,[object Object],[object Object],[object Object]

Alternative Therapeutic Methods ,[object Object],[object Object],[object Object],[object Object],[object Object]

Major Histocompatablility Complex

Other Alternative Methods ,[object Object],[object Object]

The Heart ,[object Object],[object Object],[object Object]

Study: Orlic et al. ,[object Object],[object Object],[object Object]

Orlic Study Results ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Figure 1(a) Magnification: 12x

Study Results, cont’d ,[object Object],[object Object],[object Object]

III. Policy Issues ,[object Object],[object Object],[object Object],[object Object],[object Object]

Stem Cell Viddeo http: //youtube .com/watch? v=mUcE1Y_bOQE

IV. A Transcriptional Logic for Nuclear Reprogramming Takahashi & Yamanaka/ Kit T. Rodolfa and Kevin Eggan Cell (2006) 126: 652-655 ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Trans Reprogramming continued ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Nuclear Reprogramming Continued ,[object Object],Figure 1: Reprogramming Differentiated Somatic Cells Individual factors insufficient to trigger embryonic reporter (Fbx15:  -geo). 24 previously identified genes: Self-renewal : Oct3 /4, Sox2 , Nanog Pluripotency: c-myc , Eras , Klf4 Q: What’s the magic formula? Results: Iterate through combinations. Narrowed pool down to 4 cDNAs (all above except Nanog).

Nuclear Reprogramming cont’d: Results & Discussion ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Generation of germline-competent induced pluripotent stem cells Keisuke Okita, Tomoko Ichisaka, & Shinya Yamanaka Nature 448 : 313-317 ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Generation of germline. . . continued ,[object Object],Construct : Enhanced GFP (EGFP)-internal ribosome entry site (IRES)-puromycin resistance (Pur R ) into 5’ UTR ,[object Object],[object Object],[object Object],~5% were were GFP positive - Cells indistinguishable form ES cells (morphology/proliferation*) Nanog iPS, Fbx15 iPS, and ES cells were then characterized.

Results ,[object Object],[object Object],Epigenetics: Methylation signature closer to ES cells SSLP (single sequence length polymorphism): Unique signature* Induction Efficiency: Nanog iPS cells: 0.001-0.03% Fbx15 iPS cells: 0.01-0.5% Differentiation in LIF & RA of Nanog iPS more closely resembles ES cells.

Discussion: ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Conclusion ,[object Object],[object Object],[object Object],[object Object],[object Object]

References: ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Stem Cells: Minireview Presentation

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