Faculty Advisor or Committee Member

David Adams, Advisor

Faculty Advisor or Committee Member

Jill Rulfs, Committee Member

Faculty Advisor or Committee Member

Bill Mackin, Committee Member

Identifier

etd-0422102-170335

Abstract

Amplification of hematopoietic stem cells (HSCs) from human cord blood has applications for a variety of cell therapy protocols. The purpose of this thesis (performed in collaboration with ViaCell, Inc.) was to analyze differential gene expression (especially related to cytokines) during the process of human HSC amplification in vitro. When applied to markers previously shown to be specific for HSC's and/or progenitor cells, the analysis validates ViaCell's cellular product. Total cellular RNA was isolated from cord blood samples at various stages of amplification and used to synthesize cDNAs as probes for hybridization arrays. mRNA candidates increased in cell populations enriched for stem cells were first identified using hybridization arrays, then confirmed by RT-PCR. Restriction mapping confirmed RT-PCR amplicons. The results identified several developmentally interesting cytokines (CD117, Jagged-2, Manic Fringe, and Notch) upregulated in stem cell enriched fractions. Analysis of one candidate previously shown to be a marker for HSCs and progenitors, CD117, was extended using Western blots to show a CD117-related protein upregulation. The observed upregulations did not contain many inflammatory cytokines, which could hinder survival of HSC grafts. The future hope for the non-CD117 candidates is as potential growth modifiers for stem cell samples isolated by clonogenic amplification.

Publisher

Worcester Polytechnic Institute

Degree Name

MS

Department

Biology & Biotechnology

Project Type

Thesis

Date Accepted

2002-04-22

Accessibility

Unrestricted

Subjects

stem cells, cytokines, Hematopoietic stem cells, Cytokines, Gene expression

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