Int J Stem Cells.  2012 Nov;5(2):96-103.

Effects of Growth Factors on Establishment and Propagation of Embryonic Stem Cells from Very Early Stage IVF Embryos and Their Characterization in Buffalo

• Puri G
• Kumar K
• Singh R
• Singh RK
• Yasotha T
• Ranjan R
• Kumar M
• Das BC
• Singh G
• Sarkar M
• Bag S

Abstract

BACKGROUND AND OBJECTIVES
Although ES cells have been derived from very early stage embryos in different species, but, so far ES cell line could be derived from early stage IVF embryos in buffalo. The present experiment was carried out to study the effects of different growth factors on attachment, formation of ES cell colonies, their extent of passaging and relative expression of pluripotency marker in these colonies in buffalo.
METHODS AND RESULTS
For this, 8~16 cell stages zona free IVF embryos were cultured with different culture condition viz. Control, Media-I: (Control+SCF), Media-II: (Control+SCF+bFGF) and Media-III: (Control+SCF+bFGF+IGF1). A total of 25 number of embryos were cultured in each medium. The efficiency (%) of blastomere attachment, % stem cell colony formation were recorded and number of passaging were evaluated in each culture condition. The results indicated that the efficiency of embryonic blastomere attachment, % stem cell colonies formation and propagation were significantly higher when medium was supplemented with growth factors viz. SCF, bFGF and IGF-1 (Media-III) than when supplemented with either SCF or SCF+bFGF. The expression of pluripotent genes viz Oct4, Nanog, FoxD3 and KLF4 were significantly higher (p<0.005) when medium was supplemented with three growth factors.
CONCLUSIONS
It can be concluded that when 8~16 cell stages zona free IVF embryos of buffalo was cultured on feeder, the % of blastomere attachment, % of ES cell colony formation and their further propagation were higher in ES cell medium supplemented with SCF+bFGF+IGF1 which may be due to high expression of pluripotent stem cell markers.

Keyword

Growth factors; Embryonic stem cell like cells; Buffalo

MeSH Terms

Blastomeres
Buffaloes
Cell Line
Embryonic Stem Cells
Embryonic Structures
Herpes Zoster
Insulin-Like Growth Factor I
Intercellular Signaling Peptides and Proteins
Pluripotent Stem Cells
Stem Cells
Insulin-Like Growth Factor I
Intercellular Signaling Peptides and Proteins