Spherical neural mass (SNM) is a mass of neural precursors that have been used to generate neuronal cells with advantages of long-term passaging capability with high yield, easy storage, and thawing. In this study, we differentiated neural retinal progenitor cells (RPCs) from human induced pluripotent stem cells (hiPSC)-derived SNMs. RPCs were differentiated from SNMs with a noggin/fibroblast growth factor-basic/Dickkopf-1/Insulin-like growth factor-1/fibroblast growth factor-9 protocol for three weeks. Human RPCs expressed eye field markers (Paired box 6) and early neural retinal markers (Ceh-10 homeodomain containing homolog), but did not photoreceptor marker (Opsin 1 short-wave-sensitive). Reverse transcription polymerase chain reaction revealed that early neural retinal markers (Mammalian achaete-scute complex homolog 1, mouse atonal homolog 5, neurogenic differentiation 1) and retinal fate markers (brain-specific homeobox/POU domain transcription factor 3B and recoverin) were upregulated, while the marker of retinal pigment epithelium (microphthalmia-associated transcription factor) only showed slight upregulation. Human RPCs were transplanted into mouse (adult 8 weeks old C57BL/6) retina. Cells transplanted into the mouse retina matured and expressed markers of mature retinal cells (Opsin 1 short-wave-sensitive) and human nuclei on immunohistochemistry three months after transplantation. Development of RPCs using SNMs may offer a fast and useful method for neural retinal cell differentiation.
Animals ; Cell Differentiation ; Humans* ; Immunohistochemistry ; Induced Pluripotent Stem Cells ; Methods ; Mice ; Neurons ; Photoreceptor Cells, Vertebrate ; Polymerase Chain Reaction ; Retina ; Retinal Pigment Epithelium ; Retinaldehyde* ; Reverse Transcription ; Stem Cells* ; Transcription Factors ; Up-Regulation

Ancient DNA analyses are widely used for evolutionary and phylogenetic study of mankind in anthropology and archeology. However, the DNA extraction from particularly poorly preserved ancient human samples is often unsuccessful in these analyses. In the present study, to improve the success rate of ancient DNA analysis, we introduced a high grade ancient DNA purification method using ion-exchange columns. We compared the success rate of ancient DNA analysis of this new method with that of the two methods that have been used for ancient DNA extraction, GENECLEAN(R) kit (Qbiogene) and Qiaquick column (Qiagen). Twelve ancient bone samples from Korea and Mongolia that are about 500 to 5,000 years old by an archeological estimation were used. As the DNA analysis methods, polymerase chain reaction (PCR) methods for the amplification of a mitochondrial DNA HV1 segment, a male sex determination marker DNA and M175 marker DNA that is used for the determination of O haplogroup of Y chromosome that is reportedly a common one in modern Korean people. The method developed in this study remarkably increased the success rate of DNA analysis compared with the other two methods. Using the GENECLEAN(R) kit, only two samples were amplifiable for the mitochondrial DNA, no samples for the male sex determination marker and M175 marker DNAs. Using the Qiaquick columns, nine samples were amplifiable for mitochondirial DNA, nine samples for male sex determination marker and six samples for M175 marker. The developed method allowed for the amplification of mitochondrial DNA from all samples, male sex determination marker from eight samples and M175 marker from eight samples. The results demonstrate that ion-exchange columns can be useful for the improved ancient DNA extraction in anthropology and archeology.
Anthropology ; Archaeology ; DNA* ; DNA, Mitochondrial ; Humans ; Korea ; Male ; Mongolia ; Polymerase Chain Reaction* ; Y Chromosome

Determination of male and female is important in anthropology, archeology and forensic science. This study was designed to compare genotype sex of improved amelogenin PCR amplication method with morphological sex of ancient human bones. Sixty human skulls which lived from the Bronze Age to twenties centuries and excavated in Uzbekistan were used in this study. Morphological sex was determined by Uzbekistan scientist, and genotype sex was determined by improved amelogenin PCR amplication developed in this study. Among 20 morphological males, 13 samples (65%) were genotypical male. Among 40 morphological females, 20 samples (50%) were genotypical male. In conclusion, morphological method might be inadequate for sex determination of ancient bones. The improved amelogenin PCR method will be useful in sex determination of ancient bones.
Amelogenin* ; Anthropology ; Archaeology ; Female ; Forensic Sciences ; Genotype* ; Humans ; Male ; Polymerase Chain Reaction* ; Skull ; Uzbekistan*

Many data from ancient human remains became useful by molecular approach for ancient human DNA. In anthropology, genetic sex is essential to understand marriage and burial patterns, differential mortality rates between sexes, and differential patterns by sex of disease, diet, status, and material possessions. This study was designed to determine genotype sex of 52 ancient human bones with well preserved skulls, and to compare with the orphological sex. Parts of femur and other bones were used as ancient bones excavated in Mongolia aged between bronze and Mongol period. Morphological sex was determined by Mongolian scientist, and genotype sex was determined by using biallelic marker RPS4Y for Y haplogroup. Of 52 genetic males, 10 samples were morphologically female. In conclusion, biallelic marker RPS4Y. PCR amplication method will be useful in sex determination of ancient bones.
Anthropology ; Burial ; Diet ; DNA ; Female ; Femur ; Genotype ; Humans* ; Male ; Marriage ; Mongolia ; Mortality ; Polymerase Chain Reaction ; Skull ; Y Chromosome