PrimeGen Biotech LLC

Santa Ana, CA, United States

PrimeGen Biotech LLC

Santa Ana, CA, United States

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Patent
Primegen Biotech LLC | Date: 2017-05-03

Compositions and methods for promoting tissue regeneration with gonad-derived stem cell side population cells are provided.


Patent
Primegen Biotech Llc | Date: 2015-06-30

Compositions and methods for promoting tissue regeneration with gonad-derived stem cell side population cells are provided.


Provided herein are methods of treatment to regenerate, restore or rejuvenate a tissue. Methods include making adult somatic and germ cells pluripotent for administration to a patient. Alternatively, created pluripotent cells may be differentiated to the desired tissue type and administered to a patient to repair or enhance the target tissue.


Izadyar F.,PrimeGen Biotech LLC | Wong J.,PrimeGen Biotech LLC | Maki C.,PrimeGen Biotech LLC | Pacchiarotti J.,PrimeGen Biotech LLC | And 11 more authors.
Human Reproduction | Year: 2011

BACKGROUND: This study was conducted to identify and characterize repopulating spermatogonial stem cells (SSCs) in the adult human testes. Methods Testes biopsies from obstructive azoospermic patients and normal segments of human testicular tissue were used. Flow cytometry, real-time PCR and immunohistochemical analysis were performed. Purified human spermatogonia were transplanted into busulfan-treated recipient mouse testes and integrated cells were detected by human nuclear protein antibody co-localized with stem cell and germ cell markers. Results Testicular biopsies collected from obstructive azoospermic men showed similar morphology and distribution of markers to the normal human testes. Flow cytometry showed distinct populations of stage-specific embryonic antigen-4 (SSEA-4), CD49f and CD90 positive cells in the adult human testes. SSEA-4 (+) cells showed high expression levels of SSC-specific genes and high levels of telomerase activity. Extensive colonization of human cells in the mouse testes indicates the presence of highly enriched populations of SSCs in the SSEA-4 (+) sorted cells. All the HNP (+) cells in the mouse testes were positive for germ cell marker dead box mRNA helicase and only half of them were dimly positive for c-kit. In addition, subpopulations of human spermatogonia that colonized mouse testes were positively stained for CD49f, GPR-125, Nanog and Oct-4 indicating the existence of population of cells among human spermatogonia with SSC and pluripotent characteristics. CONCLUSIONS This study clearly demonstrates that repopulating human SSCs have phenotypic characteristics of SSEA-4+, CD49f +, GPR-125+and c-Kit neg/low. The results have direct implications for enrichment of human spermatogonia for further culture and germ cell differentiation studies. © The Author 2011. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.


Ding Y.C.,Beckman Research Institute | Steele L.,Beckman Research Institute | Kuan C.-J.,University of California at Irvine | Greilac S.,PrimeGen Biotech LLC | Neuhausen S.L.,Beckman Research Institute
Breast Cancer Research and Treatment | Year: 2011

Male breast cancer (MBC) is an uncommon disease with a frequency of approximately one in 1000. Due to the rarity of MBC, it is understudied and its etiology is poorly understood. Our objectives are to determine the frequency of pathogenic mutations in BRCA2 and PALB2 in MBC cases and to investigate the correlations between mutation status and cancer phenotypes. Single strand conformation polymorphism analysis, direct sequencing, and multiplex ligation-dependent probe amplification were employed to screen for mutations in the BRCA2 gene, followed by direct sequencing of the PALB2 gene in BRCA2-negative MBC cases. Pathogenic BRCA2 mutations were identified in 18 of the 115 MBC cases, including four of the ten cases (40%) from breast cancer families and 14 of the 105 cases (13%) unselected for family history of breast cancer. The difference in BRCA2-mutation frequencies between cases with and without family history of breast cancer was not statistically significant (P = 0.145), suggesting that family history is not a strong predictor of carrying a mutation in males. We observed a highly significant association of carrying a pathogenic BRCA2 mutation with high tumor grade (P<0.001) and a weak association with positive lymph nodes (P < 0.02). Of the 97 BRCA2-negative MBC cases, we identified one PALB2 mutation with confirmed pathogenicity and one mutation predicted to be pathogenic, a prevalence of pathogenic PALB2-mutation of 1-2%. Based on our results and previous studies, genetic testing for BRCA2 should be recommended for any diagnosed MBC case, regardless of family history of breast cancer. © Springer Science+Business Media, LLC. 2010.


PubMed | Primegen Biotech LLC, Tabriz University of Medical Sciences, Tehran University of Medical Sciences and University of Kashan
Type: Journal Article | Journal: Andrologia | Year: 2016

Testicular cancer is the most common cancer affecting men in reproductive age, and cisplatin is one of the major helpful chemotherapeutic agents for treatment of this cancer. In addition, exposure of testes cancer cells to cisplatin could potentially eliminate tumour cells from germ cells in patients. The aim of this study was to evaluate the effect of cisplatin on viability of mouse acute lymphoblastic leukaemia cell line (EL-4) and neonatal mouse spermatogonial cells invitro. In this study, the isolated spermatogonial stem cells (SSC) and EL-4 were divided into six groups including control (received medium), sham (received DMSO in medium) and experimental groups which received different doses of cisplatin (0.5, 5, 10 and 15gml(-1) ). Cells viability was evaluated with MTT assay. The identity of the cultured cells was confirmed by the expression of specific markers. Our finding showed that viability of both SSC and EL-4 cells was reduced with the dose of 15g/ml when compared to the control group (P0.05). Also, the differences between the IC50 in doses 10 and 15g/ml at different time were significant (P0.05). The number of TUNEL-positive cells was increased, and the BAX and caspase-3 expressions were upregulated in EL4 cells for group that received an effective dose of cisplatin). In conclusion, despite the dramatic effects of cisplatin on both cells, spermatogonial stem cells could form colony in culture.


Supronowicz P.,Circle Biologics | Gill E.,Circle Biologics | Trujillo A.,Circle Biologics | Thula T.,Circle Biologics | And 3 more authors.
Tissue Engineering - Part A | Year: 2011

Background: Tissue engineering of new bone relies on the combination and application of osteoconductive, osteoinductive, and osteogenic elements. Natural scaffolds, such as demineralized bone matrix (DBM), contain collagenous networks with growth factors such as bone morphogenetic protein-2. Stem cells from readily available sources, including discarded adipose tissue, have the propensity to differentiate into bone. The present study examines a multi-component technique consisting of a novel side population of adipose stem cells cultured on DBM for tissue engineering applications. Methods: Adipose-derived side population stem cells were cultured on DBM for up to 14 days. Cell proliferation, alkaline phosphatase activity, extracellular matrix protein production, and calcium-containing mineral deposit formation were assayed. Ectopic bone formation in a rat model was also evaluated. Results: Side population stem cells attached to and proliferated on DBM while generating markers of new bone formation. When these cell/substrate composites were implanted into an ectopic model, newly formed bone was 30% greater than that of DBM alone. Conclusions: Novel populations of adipose-derived stem cells cultured on DBM compose a system that develops new bone matrix in vitro and in vivo. This strategy provides a novel approach using naturally occurring materials for bone repair in tissue engineering applications. © Mary Ann Liebert, Inc. 2011.


Patent
PrimeGen Biotech LLC | Date: 2014-02-03

Disclosed herein are methods for cryopreserving cells and tissues under clinical conditions, allowing production of viable cell products suitable for transplantation.


Patent
PrimeGen Biotech LLC | Date: 2012-11-28

Methods are provided for the isolation, characterization and propogation of germline cells stem cells from fetal and adult mammals. Additionally, isolated populations of germ line cells having different phenotypes are disclosed wherein the subpopulations are capable of forming long-term cultures of multipotent or pluripotent cells or are capable of differentiating into mature germline cells and repopulating a sterile reproductive organ. The multipotent or pluripotent germline cells are also suitable for differentiation into tissue-specific somatic cells for therapeutic purposes.


Pacchiarotti J.,PrimeGen Biotech LLC | Ramos T.,PrimeGen Biotech LLC | Howerton K.,PrimeGen Biotech LLC | Greilach S.,PrimeGen Biotech LLC | And 3 more authors.
BioMed Research International | Year: 2013

Recent work in preservation of female fertility as well as new information on the nature of spermatogonial stem cells has prompted an investigation into the possibility of an effective clinical-grade procedure for the cryopreservation of testicular cells and/or tissue. Clinical-grade reagents, validated equipment, and protocols consistent with cGTP/cGMP standards were used in developing a procedure suitable for the safe and effective cryopreservation of human testicular cells and tissues. These procedures were designed to be compliant with the relevant FDA regulations. The procedure proved to effectively cryopreserve both testicular cells and tissue. The cryopreservation of testicular tissue was comparable in most aspects we measured to the cryopreservation of isolated cells, except that the viability of the cells from cryopreserved testicular tissue was found to be significantly higher. On the other hand, cryopreservation of cells is preferred for cell analysis, quality control, and sterility testing. This study demonstrates that testicular tissue and cells from sexual reassignment patients can be successfully cryopreserved with a clinical-grade procedure and important cell populations are not only preserved but also enriched by the process. Further studies will determine whether these findings from hormone-treated patients can be generalized to other patients. © 2013 Jason Pacchiarotti et al.

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