Growth and gene expression in diploid epithelial cell lines derived from normal human parotid gland

Chopra, Dharam P.; Xue-Hu, Iris; Reddy, L. J.

doi:10.1046/j.1432-0436.1995.5830241.x

Cited by 6 publications

(13 citation statements)

References 40 publications

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“…Organ culture techniques, however, have limited appeal in biological applications because these do not allow studies on specifi c epithelial lineage differentiation and carcinogenesis independent of other cells. Using an explant outgrowth procedure, epithelial cell populations from several human tissues (eg, prostate, bronchus, and salivary gland) have been generated [38][39][40]. Small tissue explants are held in a dish by a plasma clot and cultured in a suitable medium.…”

Section: Primary and Long-term Intestinal Epithelial Cell Culturementioning

confidence: 99%

Intestinal Epithelial Cells In Vitro

Chopra

Dombkowski

Stemmer

et al. 2010

Stem Cells and Development

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Recent advances in the biology of stem cells has resulted in signifi cant interest in the development of normal epithelial cell lines from the intestinal mucosa, both to exploit the therapeutic potential of stem cells in tissue regeneration and to develop treatment models of degenerative disorders of the digestive tract. However, the diffi culty of propagating cell lines of normal intestinal epithelium has impeded research into the molecular mechanisms underlying differentiation of stem/progenitor cells into the various intestinal lineages. Several short-term organ/organoid and epithelial cell culture models have been described. There is a dearth of long-term epithelial and/or stem cell cultures of intestine. With an expanding role of stem cells in the treatment of degenerative disorders, there is a critical need for additional efforts to develop in vitro models of stem/progenitor epithelial cells of intestine. The objective of this review is to recapitulate the current status of technologies and knowledge for in vitro propagation of intestinal epithelial cells, markers of the intestinal stem cells, and gene and protein expression profi les of the intestinal cellular differentiation.

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Section: Primary and Long-term Intestinal Epithelial Cell Culturementioning

confidence: 99%

Intestinal Epithelial Cells In Vitro

Chopra

Dombkowski

Stemmer

et al. 2010

Stem Cells and Development

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“…More importantly the serially passaged MCF-10 cell cultures, derived from human breast epithelium, has been characterized as diploid and normal Soule et al, 1990. Recently, we have described two diploid epithelial acinar cell lines, HPAM1 and HPAF1, derived from the normal human salivary gland (Chopra and Xue-Hu, 1993;Chopra et al, 1995). The HPAMl cell line has been passaged more than 50 times (> 189 population doublings) and HPAFl more than 40 times (> 185 population doublings).…”

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confidence: 99%

Differential growth factor responses of epithelial cell cultures derived from normal human prostate, benign prostatic hyperplasia, and primary prostate carcinoma

et al. 1996

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Because of a lack of information of the optimum nutritional requirements, epithelial cells derived from normal human prostate and prostate tumors have been difficult to propagate in vitro, which hinders research in prostate carcinogenesis. In an effort to establish optimum nutritional conditions and differences in growth characteristics of normal human prostate (NP), benign prostatic hyperplasia (BPH), and prostatic carcinoma (PCA), we have compared the effects of several growth factors on cell proliferation and elucidated growth properties of low passage epithelial cells derived from NP, BPH, and PCA of an African-American patient. Primary and low passage cultures were propagated in serum-free keratinocyte basal medium (KBM) supplemented with insulin (5 micrograms/ml), hydrocortisone (0.5 microgram/ml), epidermal growth factor (EGF, 10 ng/ml), bovine pituitary extract (BPE; 50 micrograms/ml), cholera toxin (10 ng/ml), and antibiotics. Almost all NP, BPH, and PCA cells were positive for cytokeratins and prostate-specific antigen (PSA). The NP, BPH, and PCA cells were essentially diploid and lacked mutations in c-K-ras and c-Ha-ras oncogenes, and p53 tumor suppressor gene. However, they exhibited progressively accelerating growth parameters. The population doubling times of NP, BPH and PCA were 51 hr, 37 hr, and 29 hr, respectively; their saturation densities were 2.9 x 10(4)/cm2, 3.3 x 10(4)/cm2, and 7.2 x 10(4)/cm2, respectively. The NP and BPH cells required all of the growth factors in the medium, as deletion of any one of the above factors strongly inhibited their growth. The PCA cells, however, were independent of EGF and hydrocortisone. PC-3, an established human prostate cancer cell line, was independent of the growth factors tested. Fetal bovine serum (FBS) inhibited the growth of NP, BPH and PCA cells. In contrast, FBS stimulated the growth of the PC-3 cells in a concentration-dependent manner. These results indicate that in the absence of any apparent karyotype alterations and mutations in c-K-ras, c-Ha-ras and p53 genes, epithelial cells derived from NP, BPH, and PCA exhibit significant differences in their growth properties and responses to growth factors. These variations may represent early changes involved in prostate cancer, while gene mutations and cytogenetic alterations occur in advanced and/or metastatic tumors.

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“…The primers for a -amylase 1 (59 -atctagaggctgggaa-3 9 and 59 -gtcagtgtcctttcca-3 9 ) and basic proline-rich protein (59 -caagaaggcaacaatc ct-39 and 59 -ctggggaggtctggaa gg-39 ) were designed to amplify 201-bp and 135-bp products. (8) Reverse transcriptase-polymerase chain reaction Total cellular RNA was extracted from homogenized parotid tissue by the sequential addition of RNAzol ™ B (Tel-Test, Inc., Friendsw ood, TX) (0.4 ml/1 3 10 6 BEC) and chloroform (0.2 ml/2.0 ml hom ogenate). The suspension was vortexed, placed on ice for 5 min, and centrifuged at 12,000 3 g for 15 min at 4°C.…”

Section: Characterization Of Parotid Cell Culturesmentioning

confidence: 99%

Interferon-alpha Upregulates Gene Expression of Aquaporin-5 in Human Parotid Glands

Smith¹,

Siddiqui²,

Modica³

et al. 1999

Journal of Interferon & Cytokine Research

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Aquaporins are a family of homologous membrane proteins that function as highly selective water channels. Aquaporin-5 (AQP5) is uniquely present in lacrimal and salivary glands, where it accounts for normal tear and saliva production. We tested the hypothesis that orally administered human interferon-alpha (HuIFN-alpha) benefits persons with xerostomia by augmenting the production of AQP5 protein by parotid gland epithelium. Cells from three human parotid glands were cultured with and without human lymphoblastoid IFN-alpha, and assayed for AQP5 mRNA levels by reverse transcriptase polymerase chain reaction (RT-PCR), and AQP5 protein levels by Western blot. Intracellular localization of AQP5 protein was done using confocal microscopy. The functional integrity of the glandular tissue was confirmed by RT-PCR analysis of alpha-amylase 1 and basic proline-rich protein transcripts. AQP5 was constitutively expressed in human parotid gland tissue, with AQP5 protein restricted to the plasma membranes and cytoplasmic vesicles of acinar cells. IFN-alpha augmented AQP5 transcription and protein production in a concentration-dependent manner, and increased the size of intensity of staining of AQP5-containing cytoplasmic vesicles in acinar cells. We conclude that IFN-alpha upregulates AQP5 gene expression in human parotid acinar cells in vitro. To our knowledge, this is the first demonstration that IFN-alpha regulates the gene expression of an aquaporin.

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Growth and gene expression in diploid epithelial cell lines derived from normal human parotid gland

Cited by 6 publications

References 40 publications

Intestinal Epithelial Cells In Vitro

Intestinal Epithelial Cells In Vitro

Differential growth factor responses of epithelial cell cultures derived from normal human prostate, benign prostatic hyperplasia, and primary prostate carcinoma

Interferon-alpha Upregulates Gene Expression of Aquaporin-5 in Human Parotid Glands

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