1. Say he is Allah
the only one.
We all need him
and
he needs no one.

2. Presenter: Ashraf Hassanpour
Cellular Therapy for
Disease
Presenter: Ashraf Hassanpour
Advisors: Dr. Vojdani

3. Introduction

4. Introduction Liver regeneration
In vertebrates, the liver is the only internal organ that can regenerate itself
Only 25% of a liver is needed to regenerate an entire organ via hepatocyte replication

5. Basic etiologies of liver injury
Infectious
Viral hepatitis (hepatotropic and opportunistic), bacterial, fungal, parasitic
Immune mediated
Autoimmune hepatitis, GVHDa (Graft versus host disease), primary biliary cirrhosis
Drug and toxin induced
Mushroom ingestion, idiosyncratic drug reaction, acetaminophen, suicide attempt
Genetic and metabolic
Inherited metabolic diseases, fatty liver disease
Miscellaneous
Obstructive cholestasis, vascular disorders, heat stroke, life style and habits (alcoholism, fast foods, insufficient physical activity, stress), neoplasms (primary and secondary)

6. Cirrhosis Non-alcoholic fatty liver disease (NAFLD)
Nonalcoholic Steatohepatitis (NASH)

7. Cell- based therapies

8. Rationalization for cell-based therapies
Improvements in hepatic inflammatory and fibrotic microenvironments
Replenish functional hepatocytes

9. Cell sources for liver disease therapy
Hepatocytes and intrahepatic stem cells
Primary hepatocytes
Liver stem/progenitor cells
Circulating stem cells
Bone marrow - derived stem cells
Hematopoietic stem cells (HSCs) CD34 and CD133 positive
Mesenchymal stem cells (MSCs) that lack a well-defined surface antigen expression pattern
Embryonic stem cells
Induced pluripotent stem cells (iPSCs)

10. Primary hepatocytes Limited in number & Variable in quality
Not able to be expanded in vitro
The ability of hepatocytes to effectively repopulate a diseased liver appears to be limited to a select group of disorders (such as hereditary tyrosinemia, wilson disease, or progressive familial intrahepatic cholestasis)
Require immunosuppression (insufficient experience to define the amount and duration of immunosuppression needed in this setting.)
Lastly, how long these hepatocytes will be viable and the nature of their interaction with native hepatocytes remain unclear
Several approaches are in development, including hepatocytes derived from cell lines, xenotransplant of animal-derived hepatocytes, and even in vivo expansion of human hepatocytes in fumarylacetoacetate hydrolase-deficient animal incubators.

11. Liver stem/progenitor cells (LSPCs)
Liver stem/progenitor cells (also known as oval cells in rodents) are thought to represent tissue specific, bipotential precursors to liver parenchymal cells
The true ability of LSPCs to transdifferentiate into mature hepatocytes are not entirely clear.
Transplant of LSPCs has been accomplished via intrasplenic injection or infusion into a peripheral vein or the portal vein.
Require immunosuppression
Severe fibrogenic response that is driven by the activation of the hepatic progenitor compartment
Directed differentiation techniques have also allowed generation of hepatic progenitor cells from human embryonic stem cells
Adult LSPCs are available only in limited numbers, and there are ethical constraints on the use of human fetal LSPCs.

12. Circulating stem cells
Bone marrow - derived stem cells
Hematopoietic stem cells (HSCs) CD34 and CD133 positive
Mesenchymal stem cells (MSCs) that lack a well-defined surface antigen expression pattern

13. Bone marrowe - derived stem cells
True pluripotent stem cells present in bone marrow are estimated to be less than 0.1% of CD133+ cells.
The migration of these stem cells appears to be mediated by a chemoattractant, such as stromal cell-derived factor; Subsequently, the secretion of interleukin 8, matrix metalloproteinase 9, hepatocyte growth factor, and stem cell factors facilitates homing and engraftment of MSCs in the liver.

14. Bone marrow - derived stem cells
Possible mechanisms
Transdifferentiation into hepatocytes
Stimulation of native hepatocyte proliferation
Immunomodulatory effects
Cell plasticity
An antifibrotic effect
Hepatic stellate cells and myofibroblasts may derive from bone marrow stem cells
MSCs can undergo malignant transformation
The approaches for clinical use
infusion of collected autologous stem cells
mobilization of bone marrow stem cells by the administration of granulocyte colony-stimulating factor (GC-SF).

15. Clinical trials with autologous bone marrow-derived stem cells
Trials of Mesenchymal Stem Cell Transplant in Patients With Chronic Liver Diseases
Reference
Cell therapy
Dose, route
No. of patients
Type of study
Results
Amer et al, 2011
BM-MSC (bone marrowe
derived hepatocytes)
Single dose,
intrahepatic,
intrasplenic
10 Intrahepatic
 10 Intrasplenic
 20 Controls
Controlled trial
Less ascites/edema, increased albumin
Zhang et al, 2012
UC-MSC
Multiple doses,
peripheral vein
30 Treatment
 15 Controls
Randomized
controlled trial
Less ascites, decreased
MELD
Mohamadnejad
et al, 2007
BM-MSC  4
Uncontrolled trial
Decreased MELD in 2 of
4 patients
Kharaziha et al, 2009
portal vein 8
Decreased MELD
Peng et al, 2011
hepatic artery
53 Treatment
 105 Controls
Decreased T Bil, improved
INR and MELD score
et al, 2013
15 Treatment
12 Placebo
No differences between
the groups
BM-MSC ¼ bone marrowederived mesenchymal stem cells; INR ¼ international normalized ratio; MELD ¼ model of end-stage liver disease; T Bil ¼ total bilirubin; UC-MSC ¼ umbilical cord-derived mesenchymal stem cell. 15

16. Clinical trials with autologous bone marrow-derived stem cells
Trials of Sorted Hematopoietic Stem Cell Transplant in Patients With Chronic Liver Diseases
Reference
Cell therapy
Dose, route
No. of patients
Type of study
Results
Gordan et al, 2006
CD34+
Single dose, portal vein or hepatic artery 5
Uncontrolled trial
Serum albumin improved, T Bil improved
Mohamadnejad
et al, 2007
Single dose, hepatic artery 4
Serum albumin, INR, T Bil improved
Pai et al, 2008 9
CP score improved, T Bil decreased
Levicar et al, 2008
Improved T Bil
CP ¼ Child-Pugh; INR ¼ international normalized ratio; T Bil ¼ total bilirubin. 16

17. Clinical trials with autologous bone marrow-derived stem cells
Trials of GC-SF-Mobilized Hematopoietic Stem Cell Transplant in Patients With Chronic Liver Diseases
Reference
Cell therapy
Dose, route
No. of patients
Type of study
Results
Yannaki et al, 2006
GC-SF/PBMNC
Single dose,
hepatic artery 2
Uncontrolled trial
CP score improved, MELD score
improved
Gaia et al, 2006
GC-SF
Multiple doses
of GC-SF 8
Feasibility, safety study
Yan et al, 2007
GC-SF/HGF
Unclear
PBMNCs were transformed in
hepatocyte-like cells
Khan et al, 2007
GC-SF/CD34+ 4
Improved serum albumin, T Bil,
ALT
Han et al, 2008
20 GC-SF plus
PBMNC infusion
20 GC-SF
Randomized
controlled trial
GC-SF plus PBMNC group had
better liver test results
Garg et al, 2012
23 Treatment
24 Placebo
Randomized, blinded,
Improved MELD score, better
patient survival
ALT ¼ alanine aminotransferase; CP ¼ Child-Pugh; GC-SF ¼ granulocyte colony-stimulating factor; HGF ¼ hepatocyte growth factor; MELD ¼ model of end-stage liver disease; PBMNC ¼ peripheral blood mononuclear cells; T Bil ¼ total bilirubin. 17

18. Embryonic stem cells Direct differentiation technique
The major activity of hepatocyte-like cells derived from hpSC – to restore to an OTCD patient’s missing liver function, specifically functioning urea cycle.
Hepatocyte-like cells derived from hpSC could become a valuable source of cells for transplantation therapy of metabolic liver desorders caused by genetic defect:
The direct differentiation procedure allow the derivation of unlimited numbers of high-purity hepatocyte-like cells from hpSC.
Differentiated derivatives of hpSC may overcome limitations brought by histocompatibility issues because one hpSC line can be HLA matched with significant segments of the human population.
Autologous cells cannot be used for treatment OTCD patients because they carry the same genetic defect, allogenetic HLA-matched hpSC-derived cells may overcome this “genetic defect” issue.
The direct differentiation procedure allows the derivation of immature cells (progenitors) that have several advantages compared to adult liver cells: an increased proliferative capacity and plasticity; less immunogenicity; potentially superior adaptation and integration capacity; and a greater resistance to cryopreservation and ischemia.
Direct differentiation technique
ESCs that have been exposed to activin A and Wnt for 3 days formed DE. Then, FGFs and BMPs were added to cells for 5 days. Early hepatocytes were exposed to hepatocyte growth factor, dexamethasone and oncostatin M for 10–15 days for additional maturation. ESC-derived hepatocyt-like cells have been transplanted in animal models with improvement in hepatic function.

19. Ethical problems and immunologic rejection are main limiting factors as well as their potential to be teratogenic

20. Somatic cell nuclear transfer technology20

21. Induced pluripotent stem cells (iPSCs)
Retroviral transduction of Oct4, Sox2, Klf4, and c-Myc genes
(Nanog, Lin28) 21

22. Induced pluripotent stem cells (iPSCs)
Mouse and human iPSCs are highly similar to their respective embryo-derived SCs counterparts in morphology, molecular and phenotype aspects.
In human iPSCs the evidence of functional differentiation into specialized cell lineages of all three embryonic germ layers has been demonstrated.
It has been shown that HLCs could also be generated from human iPSCs.
The differentiated HLCs showed several similarities in morphology, the expression of a set of proteins, such as a-fetoprotein and albumin, and functionality such as glycogen synthesis, detoxification and engraftment, after transplantation into a suitable animal model
IT has been shown that HLCs derived from human ESCs and human iPSc exhibited broad similarity as well as meaningful differences.
Retroviral transduction of Oct4, Sox2, Klf4, and c-Myc genes 22

23. How have iPSCs been used so far?
Regenerative medicine
Disease modelling
Drug screening
Recently, several liver-specific disease iPSCs, such as familial hypercholesterolemia, glycogen storage diseases, familial hypercholesterolemia, alpha 1-antitrypsin deficiency and Crigler Najjar syndrome have been launched.

24. How have iPSCs been used so far?
Disease modelling
Regenerative medicine 24

25. Challenging issues in front of iPSCs
The somatic cell reprogrammed stem cells may potentially solve the ethic issues and the xenograft rejection problems caused by human ESCs derived cells for cell therapy, however, the traditional way of introducing reprogramming factors into somatic cells via lentivirus or retrovirus may bring another risk---tumor formation [8]. In order to overcome this problem, researchers developed several optimized approaches to reduce the risk of mutating genome, which include using plasmid DNA, RNA, RNA virus, and proteins [9-12]. In theory, after long-term passage or cellular metabolism, the induced exogenous factors could be completely removed from the cells; however, these methods are still not the best way to guarantee the safety of the introduced factors.
It has reported recently that transplantation of undifferentiated iPSCs demonstrated T-cell-dependent immune response in recipient syngeneic mice due to the abnormal expression of antigens following genetic manipulation.
Teratogenicity and their attitude toward malignancy

26. Stimulus –Triggered Acquisition of pluripotency (STAP)
Unfortunately, it looks this method has a hard time to convert adult somatic cells 26

28. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

29. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

30. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

31. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

32. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

33. Thanks for Your Attention
Wipe transitions with pictures and captions
(Basic)
To reproduce the picture effects on this slide, do the following:
On the Home tab, in the Slides group, click Layout, and then click Blank.
On the Insert tab, in the Images group, click Picture. In the Insert Picture dialog box, select a picture and then click Insert.
Select the picture. Under Picture Tools, on the Format tab, in the Size group, click the Size and Position dialog box launcher. In the Format Picture dialog box, resize or crop the image so that the height is set to 4” and the width is set to 10”. To crop the picture, click Crop in the left pane, and in the right pane, under Crop position, enter values into the Height, Width, Left, and Top boxes. To resize the picture, click Size in the left pane, and in the right pane, under Size and rotate, enter values into the Height and Width boxes.
Also in the Format Picture dialog box, click Shadow in the left pane, in the Shadow pane, click the button next to Presets, and then under Inner click Inside Top.
Also in the Format Picture dialog box, click Glow and Soft Edges in the left pane, in the Glow and Soft Edges pane, under Glow, do the following:
Click the button next to Color, click More Colors, and then in the Colors dialog box, on the Custom tab, enter values for Red: 157, Green: 177, and Blue: 91.
In the Size box, enter 8 pt.
In the Transparency box, enter 60%.
To reproduce the text effects on this slide, do the following:
On the Insert tab, in the Text group, click Text Box, and then on the slide drag to draw your text box.
Enter text in the text box, and then select the text. On the Home tab, in the Font group, do the following:
In the Font list, select Calibri.
In the Font Size list, select 24 pt.
Click the Font Color, and then under Theme Colors click White, Background 1 (first row).
Click Italic.
Position the text box below the picture.
To reproduce the transition effects on this slide, do the following:
On the Transitions tab, in the Transition to This Slide group, click More, and then click Wipe.
Also on the Transitions tab, in the Transition to This Slide, click Effect Options, and then click From Right.
Also on the Transitions tab, in the Timing group, do the following:
In the Duration box, enter 2.00 seconds.
Clear the On Mouse Click box.
Select After, and then in the After box enter 3.00 seconds.
To reproduce the second, third, and fourth slides, do the following:
In the Slides pane, select the slide. On the Home tab, in the Slides group, click the arrow below New Slide, and then click Duplicate Selected Slides. Repeat this process until there are three slides.
On the slide, select the picture. Under Picture Tools, on the Format tab, in the Adjust group, click Change Picture. In the Insert Picture dialog box, select a picture and then click Insert.
In the Slides pane, select the third slide.
To reproduce the background on this slide, do the following:
On the Design tab, in the Background group, click Background Styles, and then click Style 12 (third row).
(Note: Selecting this background changes the colors on the slide.)
Thanks for Your Attention

35. Hepatocytes and intrahepatic stem cells
Liver stem/progenitor cells (LSPCs)
Transplant of LSPCs from various sources has been accomplished via intrasplenic injection or infusion into a peripheral vein or the portal vein.
Generally, a regenerative stimulus such as partial hepatectomy or retrorsine injection is required for optimal engraftment
Severe fibrogenic response that is, in fact, driven by the activation of the hepatic progenitor compartmen
Require immunosuppression (insufficient experience to define the amount and duration of immunosuppression needed in this setting.)
The true ability of LSPCs to transdifferentiate into mature hepatocytes and the extent to which this process might contribute to liver regeneration and repair in various disease states are not entirely clear.
Directed differentiation techniques have also allowed generation of hepatic progenitor cells from human embryonic stem cells
Adult LSPCs are available only in limited numbers, and there are ethical constraints on the use of human fetal LSPCs.

37. Liver stem/progenitor cells (LSPCs)
Liver stem/progenitor cells (also known as oval cells in rodents) are thought to represent tissue specific, bipotential precursors to liver parenchymal cells
Transplant of LSPCs has been accomplished via intrasplenic injection or infusion into a peripheral vein or the portal vein.
Require immunosuppression
Severe fibrogenic response that isdriven by the activation of the hepatic progenitor compartmen
The true ability of LSPCs to transdifferentiate into mature hepatocytes are not entirely clear.
Directed differentiation techniques have also allowed generation of hepatic progenitor cells from human embryonic stem cells
Adult LSPCs are available only in limited numbers, and there are ethical constraints on the use of human fetal LSPCs.

39. Clinical trials with autologous bone marrow-derived stem cells
Trials of Unsorted Bone Marrow-Derived Mononuclear Cell Transplant in Patients With Chronic Liver Diseases
Reference
Cell therapy
Dose, route
No. of patients
Type of study
Results
Lyran et al, 2007
BM-MNC
Single dose,
hepatic artery 10
Uncontrolled trial
Decreased T Bil and INR, increased
serum albumin
 Terai et al, 2006
peripheral vein 9
Improved serum albumin, total protein,
CP score
 Kim et al, 2010
Less ascites, improved CP scores,
increased liver volume
 Saito et al, 2011
5 Treatment
5 Controls
Randomized
controlled trial
Improved CP scores and INR, higher
serum albumin and total protein
 Lyra et al, 2010
15 Treatment
15 Controls
Improved serum albumin and CP score
Spahr et al, 2013
BM-MNC +
GC-SF
28 Treatment
30 Controls
No significant differences between
study groups
BM-MNC ¼ bone marrowederived mononuclear cells; CP ¼ Child-Pugh; GC-SF ¼ granulocyte colony-stimulating factor; INR ¼ international normalized ratio; T Bil ¼ total bilirubin. 39