# DIFFERENTIATION OF NEURAL CREST CELLS IN VITRO

> **NIH NIH R01** · MEDICAL COLLEGE OF WISCONSIN · 1994 · $189,084

## Abstract

The long term objective of the proposed work is to elucidate some of the
mechanisms that govern commitment of pluripotent cells to a particular
cell lineage.  Quail and mouse neural crest cells will be used as an
experimental system.  the neural crest originates in the neural folds and
gives rise to a wide variety of cell types in the mature vertebrate
organism.  They include the peripheral nervous system, most primary
sensory neurons, endocrine cells, pigment cells, and the cranial
ectomesenchymal derivatives, muscle, bone, cartilage, and dermis.  The
majority of neural crest cells are pluripotent as they leave the neural
tube.  Moreover, pluripotent cells are also present at final sites of
neural crest cell differentiation, supporting the notion of location-
specific cues that originate from the embryonic microenvironment and
influence neural crest cell differentiation.  since up to 9 different
cell types are generated in neural crest cell cultures, this is a
valuable experimental system to investigate the mechanisms that govern
differentiation of pluripotent cells into a wide spectrum of phenotypes.
The proposed work is presented in three specific aims.  1) The influence
of the four growth factors, neurotrophin-3 (NT-3), basic fibroblast
growth factor (bFGF), transforming growth factor-beta (TGF-beta), and
stem cell factor (SCF) will be investigated by in vitro clonal analysis
of quail neural crest cells.  The aim is to perform these studies in a
define culture medium.  Emphasis is placed on discriminating between
trophic actions that affect cell growth/survival and influences that are
involved in cell commitment.  2) The newly developed in vitro clonal
culture system for pluripotent mouse neural crest cell swill be used to
analyze differentiation of neural crest cells derived from normal
C57BL/6J mice and the mutant strains White spotting (W), Steel (Sl), and
Piebald (s).  3) Putative novel regulatory genes involved in cell type
specification will be searched for by PCR-amplification of qual RNA
obtained from a critical stage in development.  to this end, pools of
degenerate primers recognizing conserved regions of sequences encoding
known DNA-binding proteins will be used.  Primary emphasis is placed on
helix-loop-helix (HLH) protein-encoding messages, because HLH proteins
are known to be involved in cell type specification.  However, primers
will also be designed to investigate zinc finger sequences, the homeotic
gene Hox-7, and the paired box gene , Pax-3.  Homologues of interest will
be characterized, sequenced, cloned into Bluescript, and subsequently
used as probes to screen three existing libraries.  The neural crest is
of clinical relevance because there are many human neural crest-derived
diseases.  They include genetic birth defects and malformations caused
by teratogens, such as various craniofacial malformations (e.g. cleft
lip/cleft palate), congenital defects of the cardiac outflow tract, and
pigmentation disorders (e.g., albini...

## Key facts

- **NIH application ID:** 2198281
- **Project number:** 5R01HD021423-09
- **Recipient organization:** MEDICAL COLLEGE OF WISCONSIN
- **SAM.gov UEI:** E8VWJXMMUQ67
- **Principal Investigator:** MAYA F SIEBER-BLUM
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $189,084
- **Award type:** 5
- **Project period:** 1985-08-01 → 1996-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2198281

## Citation

> US National Institutes of Health, RePORTER application 2198281, DIFFERENTIATION OF NEURAL CREST CELLS IN VITRO (5R01HD021423-09). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2198281. Licensed CC0.

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