The document summarizes a study that found that the adhesive properties and phenotypes of primary cells isolated from rats were altered if the rats' mothers were fed a low protein diet during pregnancy and nursing. Specifically, primary cells from rats exposed to maternal undernutrition showed slower adhesion and were less able to adhere to micropatterns than cells from unexposed rats, even when the exposed rats received tryptophan supplementation. This suggests that maternal diet can have long-lasting effects on offspring cell physiology that are retained in cell cultures established from biopsies.

1. The adhesive properties
of tail Primary cells tell a
tale about mother’s diet
Figure1.Morphologyofrepresentativeprimarycells,allexpressing
tryptophan hydroxylase (red) on CYTOO Starter micropatterns.
Similar diversity of phenotypes was found on primary cells
isolated from rat submitted to prenatal undernutrition or not.
Supplementation with a perinatal bolus of L-tryptophan did not
alter the phenotypes. © Plos One 2013 Nascimento et al.
“Our results suggest that adhesion
mechanisms are deeply altered in
rats raised from a mother fed on
low protein diet. ”
Bertrand Kaeffer, PhD, UMR-
1280, Nantes, France
Elizabeth Nascimento, from UMR-1280 in Nantes, France, has
recently published with colleagues a paper in Plos One showing that
the non-invasive recovery of fibroblasts from rats can be used to evaluate
the consequences of mother’s low protein diet and of perinatal tryptophan
supplementation on cell physiology. It is known that early-life stress induces
metabolicadaptationsthatinitiallyfavorsurvivalbutareultimatelydetrimentaltoadult
health.This study brings new insight on the long-term effects of early-life undernutrition.
Hush little baby, no pain, no cry: tiny biopsies !
To investigate the capacity of colonization at seeding, the adhesion properties and phenotypes of primary cells,
Nascimiento inoculated freshly trypsinized primary cells from small biopsies of the rats’ tail on conventional 2D
culture plastic.A week later,primary cultures with actively growing cellular colonies were enumerated to calculate
the capacity of colonization at seeding, and subcultured
to establish cell lines. Cells from 4 representative
fibroblast cell lines of Low Protein mother diet and
Control groups were then inoculated on CYTOO Starter
chips. The different 2D+ micropatterns were used to
test the homogeneity of cellular preparations and their
capacity to adhere to a substratum under a controlled
microenvironment (Figure 1).
Primary cells from biopsies of young rats showed
a capacity of colonization of 100%. However,
Nascimiento found that the speed of adhesion
was slower with cell preparations from the
«Low-Protein» rats. The diversity of cell
phenotypes as well as the expression of
tryptophan-hydroxylase were identical
between established cell lines from both
groups.

2. Further reading:
Nascimento E, Guzman-Quevedo O, Delacourt N, et al. Long-Lasting Effect of Perinatal Exposure to L-tryptophan on Circadian Clock of Primary
Cell Lines Established from Male Offspring Born from Mothers Fed on Dietary Protein Restriction. PLoS ONE. 2013;8(2):e56231.
The number of cells able to adhere
to fibronectin micropatterns from the
«Low Protein» group (receiving a daily bolus
of L-tryptophan or of saline solution) was lower
(percentage of occupied micropatterns with a cell: 23.6%
±7.6%) than for the cells isolated from rats of the Control group
(percentage of occupied micropatterns with a cell: 65.9% ±4.9%)
confirming observation at the onset of primary cultures. Primary cells
collected from adult rats were significantly less able to colonize plastics after
one week in culture than corresponding cells from young rats, and mothers’ Low
Protein Diet affected their property ever further.
The capacity of colonization at seeding and the adhesion potential of primary cells were
clearly altered in rats born from mothers fed on dietary protein restriction, irrespectively of the
supplementation with L-tryptophan. Primary cells retained nutritional stress-related alterations, a
source of functional assays on living cells.
Are you sleeping, are you sleeping? Brother John,
Brother John?
From human studies, it is known that the circadian
rhythm of tryptophan in breast milk affects the rhythms of
6-sulfatoxymelatonin and sleep in newborn and that infant
formulas supplemented in L-tryptophan during the night can
alter the expression of genes in cerebellum of nursing rat
neonates. Nascimiento’s results indicate that milk formulas
designed to improve sleep-wake cycles of babies can be
tested on rat models under several conditions of feeding
to check for global phenotypic consequences.Functional
assays can be designed with living cells which may
be sampled by non-invasive means in long term
experiments in epidemiological surveys.
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Figure 2: The Starter’s CYTOOchip layout: 4 different
patterns on the same chip and 3 sizes. Identical
patterns are grouped in zones of 3 x 4 blocks.
Few primary cells tell a lot on
your first bottle of milk !