Establishment and characteristics of embryonic stem cells lines from Kunming mice

An XIE; Yang WANG; Yuanlei LOU; Lixing ZHANG; Nianhua FENG

ABSTRACT

BACKGROUND:

Up to date,inbred embryonic stem cells(ESCs) line mainly derived from 129 mouse strain and C57BL/6 strain,occasionally from BALB/c mouse strain,however,few reports concerning ESCs lines derived from Ch inese Kunmingmouse strain.

OBJECTIVE:

To isolate and culture mouse ESCs of kunming species,additionally,to identify its biological properties.DESIGN,TIME AND

SETTING:

The experiment with cells as observed subjects was conducted in the institute of Urology,First Affiliated Hospital of Nanchang University from May 2006 to June 2007.MATERIALSBlastocysts of Kunmin mice with 3.5 days of embryonic age.

METHODS:

Inner cell mass from 3.5 days embryonic age were isolated from Kunming species mice,cultured on feeder layers of primary mice embryonic fibroblasts,and then the cells were isolated and subsequently cultured.MAIN OUTCOME

MEASURES:

Colony growth was observed and determined by alkaline phosphatase (AKP) staining;The expression of stage-specific embryonic antigen (SSEA)-1,and cell specific genes of c-Myc,Nanog,Oct3/4 and Sox2 were measured by immunofluorescence and RT-PCR;finally,the ability of differentiation in vitro and in vivo was identified.

RESULTS:

The ESCs-like colonies presented typical morphological characteristics of ESCs,which was positive to AKP and SSEA-1,and could express several cell specific genes,such as c-Myc,Nanog,Oct3/4 and Sox2,and differentiate into various cell types in vitro and in vivo.

CONCLUSION:

An ESCs line is successfully dedved from Kunming mice,which has typical biological characteristics of ESCs.