Models for our project

Introduction

Blalalala

Cell Lines

Throughout our project we worked with 4 mammalian cell lines:

HEK293

Human embryonic kidney cells 293

HEK293 are a widely used adherent cell line, derived from human embryonic kidney cells, that was transformed by human adenovirus 5 DNA. ^[1] We chose these cells to be one of our two model cell lines because of they are easy to grow in culture and highly susceptible to chemical transfection.

The cells were kindly provided by Prof. Roee Amit's lab

HPC-7

HPC-7 cells are immortalized murine hematopoietic progenitor cells. This cell line was created by transfecting murine embryonic stem cells with the LHx2 gene.^[2] The differentiating potential of these cells is unclear. While they are considered multipotent^[3]^[4], it appears that they are actually common myeloid progenitors according to a paper from 1998.^[5]

The cells are grown in suspension, and are very fragile and difficult to grow and transfect.

HPC-7 cells were chosen as our second model cell line because they display many characteristics of hematopoietic stem cells ^[2] and serve as a recognized model for HSCs ^[3]. The cells were kindly given to us by Professor Ben-Zion Levi's Lab

WEHI 231

WEHI 231 cells are a murine immature B cell lymphoma line. These suspension cells express the phenotype of immature B cells and display IgM antibodies on their membrane. These features made them perfect for our proof of concept experiment . The cells were kindly provided by Doron Melamed's lab .

CHO-SCF

Chinese hamster ovary cells are epithelial cells derived from a hamster's ovary.

These adherent cells are known for their ability to secrete proteins into the cell culture supernatant. We utilized this property to create stem cell factor, which needed in order to grow the HPC-7 cells. The cells were kindly given by Professor Ben-Zion Levi's Lab

Graham FL, Smiley J, Russell WC, Nairn R ,"Characteristics of a human cell line transformed by DNA from human adenovirus type 5". J. Gen. Virol. 36 (1): 59–74,1977.
Dean P. J. Kavanagh, Adrian I. Yemm, Yan Zhao, Jon Frampton, Mechanisms of Adhesion and Subsequent Actions of a Haematopoietic Stem Cell Line, HPC-7, in the Injured Murine Intestinal Microcirculation In Vivo, PLOS ONE , Volume 8 ,2013
Ross C. Hardison, Finding partners to play the music of regulation, Blood; 127(13): 1624–1626,2016.
Berthold Göttgens et al. Combinatorial Transcriptional Control In Blood Stem/Progenitor Cells: Genome-wide Analysis of Ten Major Transcriptional Regulators,Cell Stem Cell , Volume 7, Issue 4, p532–544, 8 October 2010 .
Perpetua Pinto do O , Åsa Kolterud and Leif Carlsson, Expression of the LIM-homeobox gene LH2 generates immortalized Steel factor-dependent multipotent hematopoietic precursors , The EMBO Journal Vol.17 No.19 pp.5744–5756, 1998.