PMID- 16499451
OWN - NLM
STAT- MEDLINE
DCOM- 20060516
LR  - 20061115
IS  - 1076-3279 (Print)
IS  - 1076-3279 (Linking)
VI  - 12
IP  - 1
DP  - 2006 Jan
TI  - Enhanced neurotrophin synthesis and molecular differentiation in non-transformed 
      human retinal progenitor cells cultured in a rotating bioreactor.
PG  - 141-58
AB  - One approach to the treatment of retinal diseases, such as retinitis pigmentosa, 
      is to replace diseased or degenerating cells with healthy cells. Even if all of 
      the problems associated with tissue transplant were to be resolved, the 
      availability of tissue would remain an ongoing problem. We have previously shown 
      that transformed human retinal cells can be grown in a NASA-developed 
      horizontally rotating culture vessel (bioreactor) to form three-dimensional-like 
      structures with the expression of several retinal specific proteins. In this 
      study, we have investigated growth of non-transformed human retinal progenitors 
      (retinal stem cells) in a rotating bioreactor. This rotating culture vessel 
      promotes cell-cell interaction between similar and dissimilar cells. We cultured 
      retinal progenitors (Ret 1-4) alone or as a co-culture with human retinal pigment 
      epithelial cells (RPE, D407) in this system to determine if 3D structures can be 
      generated from non-transformed progenitors. Our second goal was to determine if 
      the formation of 3D structures correlates with the upregulation of neurotrophins, 
      basic fibroblast growth factor (bFGF), transforming growth factor alpha 
      (TGFalpha), ciliary neurotrophic factor (CNTF), and brain-delivered neurotrophic 
      factor (BDNF). These factors have been implicated in progenitor cell 
      proliferation, commitment, differentiation, and survival. We also investigated 
      the expression of the following retinal specific proteins in this system: neuron 
      specific enolase (NSE); tyrosine hydroxylase (TH); D(2)D(3), D(4) receptors; 
      protein kinase-C alpha (PKCalpha), and calbindin. The 3D structures generated 
      were characterized by phase and scanning transmission electron microscopy. 
      Retinal progenitors, cultured alone or as a co-culture in the rotating 
      bioreactor, formed 3D structures with some degree of differentiation, accompanied 
      by the upregulation of bFGF, CNTF, and TGFalpha. Brain-derived neurotrophic 
      factor, which is expressed in vivo in RPE (D407), was not expressed in monolayer 
      cultures of RPE but expressed in the rotating bioreactor-cultured RPE and retinal 
      progenitors (Ret 1-4). Upregulation of neurotrophins was noted in all rotating 
      bioreactor-cultured cells. Also, upregulation of D(4) receptor, calbindin, and 
      PKCalpha was noted in the rotating bioreactor-cultured cells. We conclude that 
      non-transformed retinal progenitors can be grown in the rotating bioreactor to 
      form 3D structures with some degree of differentiation. We relied on molecular 
      and biochemical analysis to characterize differentiation in cells grown in the 
      rotating bioreactor.
FAU - Kumar, Ravindra
AU  - Kumar R
AD  - Department of Pathology, Morehouse School of Medicine, Atlanta, Georgia, USA.
FAU - Dutt, Kamla
AU  - Dutt K
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Tissue Eng
JT  - Tissue engineering
JID - 9505538
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Ciliary Neurotrophic Factor)
RN  - 0 (Nerve Growth Factors)
RN  - 0 (RNA, Messenger)
RN  - 0 (Transforming Growth Factor alpha)
RN  - 103107-01-3 (Fibroblast Growth Factor 2)
SB  - IM
MH  - *Bioreactors
MH  - Brain-Derived Neurotrophic Factor/biosynthesis/genetics
MH  - Cell Culture Techniques
MH  - Cell Differentiation/*physiology
MH  - Cell Line
MH  - Cells, Cultured
MH  - Ciliary Neurotrophic Factor/biosynthesis/genetics
MH  - Coculture Techniques
MH  - Fibroblast Growth Factor 2/biosynthesis/genetics
MH  - Humans
MH  - Microscopy, Electron, Scanning
MH  - Nerve Growth Factors/*biosynthesis
MH  - RNA, Messenger/biosynthesis
MH  - Retina/*cytology/metabolism
MH  - Stem Cells/cytology/*metabolism
MH  - Transforming Growth Factor alpha/biosynthesis/genetics
EDAT- 2006/02/28 09:00
MHDA- 2006/05/17 09:00
CRDT- 2006/02/28 09:00
PHST- 2006/02/28 09:00 [pubmed]
PHST- 2006/05/17 09:00 [medline]
PHST- 2006/02/28 09:00 [entrez]
AID - 10.1089/ten.2006.12.141 [doi]
PST - ppublish
SO  - Tissue Eng. 2006 Jan;12(1):141-58. doi: 10.1089/ten.2006.12.141.