Abstract
Previous work determined that bovine interleukin-6 (IL6) increases inner cell mass (ICM), primitive endoderm (PE), and total cell number in in vitro produced (IVP) bovine blastocysts. Another IL6 family member, leukemia inhibitory factor (LIF), has the potential to produce the same effects of IL6 due to the presence of its receptor in bovine blastocysts. We compared the abilities of LIF and IL6 to increase ICM cell numbers in day 7, 8, and 9 IVP bovine blastocysts. Supplementation with 100 ng/ml LIF from day 5 onward improved blastocyst formation rates on days 7 and 8 similar to what was observed when supplementing 100 ng/ml IL6. However, LIF supplementation did not cause an increase in ICM numbers like was observed after supplementing IL6. On day 9, increases in PE cell numbers were detected after LIF supplementation, but 300 ng/ml LIF was required to achieve the same effect on PE numbers that was observed by providing 100 ng/ml IL6. Collectively, these results show that LIF can mimic at least some of the effects of IL6 in bovine blastocyst.
Highlights
Leukemia inhibitory factor (LIF) is a well-recognized pluripotency factor in mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells, where it facilitates the maintenance of cells in a state reminiscent of inner cell mass (ICM) or epiblast (EPI) cells (Fernandes et al, 2010; Fraga et al, 2011; Fernandez-Alonso et al, 2017)
The effect of IL6 or LIF supplementation on blastocyst development and cell numbers were determined in a study where embryos were supplemented with either 100 ng/ml IL6, 100 ng/ml LIF or carrier only (1% [w/v] bovine serum albumin (BSA)) beginning at day 5 (Figure 1)
On day 7, neither LIF nor IL6 supplementation affected the percentage of cleaved embryos that formed blastocysts (Figure 1A)
Summary
Leukemia inhibitory factor (LIF) is a well-recognized pluripotency factor in mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), where it facilitates the maintenance of cells in a state reminiscent of inner cell mass (ICM) or epiblast (EPI) cells (Fernandes et al, 2010; Fraga et al, 2011; Fernandez-Alonso et al, 2017). Blastocyst development, hatching, and/or post-thaw embryo recovery were increased following LIF supplementation in some studies (Yamanaka et al, 1999; Neira et al, 2010; Kocyigit and Cevik, 2015, 2017) but not others (Fukui and Matsuyama, 1994; Sirisathien and Brackett, 2003; Vejlsted et al, 2005; Rodriguez et al, 2007). Studies have not detected improvements in ICM cell numbers after LIF exposure, but rather, two studies detected reduced ICM numbers and hypoblast development (Vejlsted et al, 2005; Rodriguez et al, 2007)
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