https://repository.kulib.kyoto-u.ac.jp/dspace/bitstream/2433/159777/1/j.cell.2006.07.024.pdf

Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors.

Human mesenchymal stem cells are thought to be multipotent cells, which are present in adult marrow, that can replicate as undifferentiated cells and that have the potential to differentiate to lineages of mesenchymal tissues, including bone, cartilage, fat, tendon, muscle, and marrow stroma. Cells that have the characteristics of human mesenchymal stem cells were isolated from marrow aspirates of volunteer donors. These cells displayed a stable phenotype and remained as a monolayer in vitro. These adult stem cells could be induced to differentiate exclusively into the adipocytic, chondrocytic, or osteocytic lineages. Individual stem cells were identified that, when expanded to colonies, retained their multilineage potential.

http://science.sciencemag.org/content/sci/284/5411/143.full.pdf

Multilineage Potential of Adult Human Mesenchymal Stem Cells

https://repository.kulib.kyoto-u.ac.jp/dspace/bitstream/2433/49782/1/Yamanaka_Cell_131_5.pdf

Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors

Somatic cell nuclear transfer allows trans-acting factors present in the mammalian oocyte to reprogram somatic cell nuclei to an undifferentiated state. We show that four factors (OCT4, SOX2, NANOG, and LIN28) are sufficient to reprogram human somatic cells to pluripotent stem cells that exhibit the essential characteristics of embryonic stem (ES) cells. These induced pluripotent human stem cells have normal karyotypes, express telomerase activity, express cell surface markers and genes that characterize human ES cells, and maintain the developmental potential to differentiate into advanced derivatives of all three primary germ layers. Such induced pluripotent human cell lines should be useful in the production of new disease models and in drug development, as well as for applications in transplantation medicine, once technical limitations (for example, mutation through viral integration) are eliminated.

https://science.sciencemag.org/content/sci/318/5858/1917.full.pdf

Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells

We report here that cells co-purifying with mesenchymal stem cells--termed here multipotent adult progenitor cells or MAPCs--differentiate, at the single cell level, not only into mesenchymal cells, but also cells with visceral mesoderm, neuroectoderm and endoderm characteristics in vitro. When injected into an early blastocyst, single MAPCs contribute to most, if not all, somatic cell types. On transplantation into a non-irradiated host, MAPCs engraft and differentiate to the haematopoietic lineage, in addition to the epithelium of liver, lung and gut. Engraftment in the haematopoietic system as well as the gastrointestinal tract is increased when MAPCs are transplanted in a minimally irradiated host. As MAPCs proliferate extensively without obvious senescence or loss of differentiation potential, they may be an ideal cell source for therapy of inherited or degenerative diseases.

/pdf/pluripotency-of-mesenchymal-stem-cells-derived-from-adult-200s4qlizf.pdf

Pluripotency of mesenchymal stem cells derived from adult marrow

Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages. After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm). These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.

Embryonic Stem Cell Lines Derived from Human Blastocysts

We have previously reported that high concentrations of basic fibroblast growth factor (bFGF) support feeder-independent growth of human embryonic stem (ES) cells, but those conditions included poorly defined serum and matrix components. Here we report feeder-independent human ES cell culture that includes protein components solely derived from recombinant sources or purified from human material. We describe the derivation of two new human ES cell lines in these defined culture conditions.

Derivation of human embryonic stem cells in defined conditions.

We prospectively evaluated 75 patients by fiber-optic bronchoscopy and bronchoalveolar lavage (BAL) for the presence of bacterial lower-respiratory-tract infection. BAL specimens were cultured quantitatively for aerobic bacteria, and a cell differential was obtained of the BAL cell population. In 18 "control" patients without evidence of respiratory infection, the presence of Mqfo squamous epithelial cells (SECs) in the BAL sample accurately predicted the presence of heavy contamination of the sample by oropharyngeal flora. In the remaining "study" patients with potential infection, polymorphonuclear leukocytes were readily identified, and potential lower-respiratory-tract pathogens were recovered in concentrations MO5 colony-forming units (cfu) per milliliter in 16 of 18 patients with bacterial infection (none had M^o SECs in their BAL sample). No patients without evidence of bacterial infection and with ^qfo SECs had MO5 cfu/ml in BAL cultures. These studies establish the ability of BAL techniques to diagnose bacterial respiratory infection.

Diagnosing Bacterial Respiratory Infection by Bronchoalveolar Lavage

As a result of their pluripotency and potential for unlimited self-renewal, human embryonic stem cells (hESCs) hold tremendous promise in regenerative medicine. An essential prerequisite for the widespread application of hESCs is the establishment of effective and efficient protocols for large-scale cell culture, storage, and distribution. At laboratory scales hESCs are cultured adherent to tissue culture plates; these culture techniques are labor-intensive and do not scale to high cell numbers. In an effort to facilitate larger scale hESC cultivation, we investigated the feasibility of culturing hESCs adherent to microcarriers. We modified the surface of Cytodex 3 microcarriers with either Matrigel or mouse embryonic fibroblasts (MEFs). hESC colonies were effectively expanded in a pluripotent, undifferentiated state on both Matrigel-coated microcarriers and microcarriers seeded with a MEF monolayer. While the hESC expansion rate on MEF-microcarriers was less than that on MEF-plates, the doubling time of hESCs on Matrigel-microcarriers was indistinguishable from that of hESCs expanded on Matrigel-coated tissue culture plates. Standard hESC cryopreservation methodologies are plagued by poor viability and high differentiation rates upon thawing. Here, we demonstrate that cryopreservation of hESCs adherent to microcarriers in cryovials provides a higher recovery of undifferentiated cells than cryopreservation of cells in suspension. Together, these results suggest that microcarrier-based stabilization and culture may facilitate hESC expansion and storage for research and therapeutic applications.

/pdf/scalable-culture-and-cryopreservation-of-human-embryonic-t5np01ynja.pdf

Scalable culture and cryopreservation of human embryonic stem cells on microcarriers.

The efficacy of bronchoalveolar lavage (BAL) in diagnosing pulmonary hemorrhage was studied in 51 immunosuppressed patients with new pulmonary infiltrates. Similar studies were performed in a control group of 8 nonimmunocompromised patients. Hemosiderin content in Prussian-blue-stained alveolar macrophages obtained by BAL was graded using a numerical scale. This “hemosiderin score” correlated closely with the degree of hemorrhage seen in corresponding histologic sections in the 26 patients from whom a lung biopsy or autopsy specimen was available. Severe pulmonary hemorrhage was ultimately diagnosed in 14 cases, and a mild degree of hemorrhage was found in an additional 19 cases. Thrombocytopenia and invasive fungal infections were statistically associated with severe hemorrhage, as was an increased percentage of alveolar macrophages in the BAL sample. This study demonstrates the efficacy of BAL in diagnosing occult pulmonary hemorrhage in the immunosuppressed host and highlights risk factors associated w...

Jeffrey M. Jones

Papers

Embryonic Stem Cell Lines Derived from Human Blastocysts

Derivation of human embryonic stem cells in defined conditions.

Diagnosing Bacterial Respiratory Infection by Bronchoalveolar Lavage

Scalable culture and cryopreservation of human embryonic stem cells on microcarriers.

Diagnosis of pulmonary hemorrhage in the immunocompromised host