JSM Cell and Developmental Biology

In vivo Transfer of Reconstructed Goat Chimeric Embryos Produced by Microinjection of Embryonic Stem Cells into Tetraploid IVF Embryos

Research Article | Open Access Volume 5 | Issue 1 |

  • 1. Department of Animal Physiology and Reproduction, ICAR-CIRG, India
+ Show More - Show Less
Corresponding Authors
Kharche SD, Department of Animal Physiology and Reproduction, ICAR-CIRG, Makhdoom P.O. Farah-28112, Mathura (UP) India, Tel: 919897987074; Fax: 915652763246

The aim of this study to observe in vivo development of a chimeric embryo (4n/2n) produced by tetraploid complementation. Cumulus oocyte complexes (COC’s, 585) were collected and matured in maturation media. Out of which 524 matured oocyes were randomly divided into two groups. Group 1(n= 296) were activated with 5μM Calcium Ionophore for 5-7 minutes followed by treatment with 2.0 mM DMAP for 4 hr in mCR2 aa medium while Group 2 (n= 228) matured oocytes were used for in vitro fertilization. Furthermore, inner cell mass (ICM) from hatched blastocysts of parthenogenetic activated embryos were used to produce ES cell-like cells while 2 cell embryos obtained from IVF were used for their production of tetraploid embryos. Chimeric embryos were produced by microinjection of parthenogenetic ESCs in to in-vitro fertilized tetraploid caprine embryos. The percentage of 2 cell, 4 cell, 8-16 cell, morula, hatched blastocyst and cleavage rate of parthenogenetic goat embryos were 20.03 ± 5.09%, 20.02 ± 6.30%, 24.41 ± 4.12%, 25.61 ± 7.63%, 9.92 ± 3.97% and 72.78 ± 13.39%, respectively while the cleavage rate following IVF was 37.08 ± 4.34%. Furthermore, 75.55 ± 12.37% embryonic stem cell colonies were formed from hatched blastocysts. The IVF two cell embryo used for tetraploid embryo production, resulting in 75.09 ± 7.86% fusion rate and 86.5 ± 13.02% cleavage rate. A total of eight microinjected chimeric embryo were transfer in two naturally synchronized goat. The result indicated that caprine ES-like cells can be incorporated into IVF tetraploid embryos by microinjection so as to produce chimeric embryos. However, in vivo development of chimeric embryos could not achieve.


Kharche SD, Pathak J, Sikarwar AS, Gangwar C, Priyadharsini R, et al. (2017) In vivo Transfer of Reconstructed Goat Chimeric Embryos Produced by Microinjection of Embryonic Stem Cells into Tetraploid IVF Embryos. JSM Cell Dev Biol 5(1): 1021.


•    Caprine
•    Chimeric embryo
•    Embryonic stem cells
•    Parthenogenetic embryo
•    Tetraploid embryo


Dairy goat is one of livestock species that provides milk and meat. It is also ideal for the transgenic production of therapeutic recombinant proteins [1]. In the last two decades there has been a great revolution in the field of biotechnology and embryo transfer technology has become a tool for enhancing lifetime productivity of livestock. In vitro embryo production is an ‘attractive alternative’ for the production of large number of embryos required for the rapid multiplication of superior germplasm of goats, upgrading a commercial or pure bred breeding programme and conservation of endangered breeds of goats [2].

Parthenogenesis is the biological phenomenon by which embryonic development is initiated without male contribution. The parthenogenetic activation of oocytes is an important tool to investigate the comparative roles of paternal and maternal genomes in controlling early embryo development [3].

Various experimental techniques have been developed over the years to maintain the full developmental potential of parthenogenetic embryos and one of the most important advances is the application of tetraploid embryos as host cells since tetraploid cells have only a limited potential in postimplantation development [4]. In earlier studies several ES cell lines were used to produce completely ES cell derived foetuses by aggregation with tetraploid morulae; however, the obtained ES foetuses died at birth [4]. Further studies demonstrated successful generation of viable and fertile ES mice which were derived exclusively from ES cells when early passage wild-type R1 [5], and TT2 [6], cells were used for aggregation with tetraploid morulae.

An alternative method, developed to reduce the damage induced by microsurgery, consisted of mechanical transfer of ESC into the embryo cytoplasm by drilling the zonapellucida (ZP).This approach includes the generation of 2n/4n chimeric embryos by microinjection of ES cells into ivf 4n embryos. Despite the present inefficiency, the generation of viable parthenogenetic goat directly from ES cells has many advantages.

In the present study we describe a successful route towards the production of parthenogenetic goat by microinjection of ES cells into tetraploid IVF embryos. However, the goal to obtain viable parthenogenetic goat using 2n/4n complementation was not reached but this technique allows viable and fertile chimeric embryos.


All organic and inorganic chemicals were purchased from Sigma Chemicals Co. except Research Vitro Cleave (RVCL) media from Cook Medical, Australia.

Parthenogenetic activation

Oocytes were collected from caprine ovaries obtained from an abattoir at Agra and were graded under the inverted phase contrast as per the method of [7]. Only grade A, B, C quality oocytes were selected and allowed to mature for 27 hr in humidified atmosphere of 5% CO2 at 38.50 C in a CO2 incubator. Maturation medium consisted of TCM-199 containing L-glutamine (100 µg/ ml), sodium pyruvate (0.25 mmol), gentamycin (50 µg/ml), FSH (5µg/ml), LH (10 µg/ml) and oestradiol-17β (1µg/ml), 10% FBS, 10% follicular fluid and 3 mg/ml BSA. To induce parthenogenetic activation, after removal of cumulus cells, denuded oocytes were activated with 5μM calcium Ionophore for 5-7 minutes followed by treatment with 2.0 mM DMAP for 4 hr in mCR2 aa medium. The embryos were then washed and cultured in embryo development medium (RVCL) followed by incubation in a CO2 incubator at 38.50 C and 5% CO2 in a humidified atmosphere. For cleavage rate, activated oocytes were observed after 48 hr under an inverted phase contrast microscope.

Embryonic stem cells production

Inner cell mass (ICM) was mechanically isolated from expanded and hatched blastocysts produced by parthenogenetic activation using microblade and were cultured on Mitomycin-C inactivated goat fetal fibroblasts feeder layer in stem cell culture medium. Passage one was performed upon primary colony formation. Subsequent colonies were passaged mechanically using a microblade every 4-5 day and media was replaced every 24 hr.

In vitro fertilization

In vitro fertilization was carried out as per the method described by [8], with slight modifications. Denuded oocytes after maturation were transferred in 50µl drops of Fert-TALP medium [9], containing 10% FBS, 8 mg/ml fatty acid free BSA and 50µg/ml heparin and the drops were inseminated with 15-20µl of the final diluted semen so as to obtain a sperm concentration of 1-2×106 sperm/ml. After in vitro insemination, the oocytes and sperm were co-incubated for 18 hr at 38.50 C with 5% CO2 in humidified atmosphere. After 18h of sperm-oocytes co-incubation, oocytes were washed and transferred in embryo development medium, Research Vitro Cleave (RVCL) media supplemented with 1% BSA for 48 hr in humidified atmosphere of 5% CO2 at 38.5ºC in CO2 incubator. The embryo development was observed under inverted phase contrast microscope up to 12 days.

Collection of two-cell embryos and electrofusion

In order to produce tetraploid embryos, the blastomeres of 2 cell diploid IVF embryos at the two-cell stage were fused after a short electric pulse. Briefly, two-cell embryos were equilibrated in 0.3 M mannitol solution for 5-10sec before they were placed individually between two platinum electrodes in 0.3 M mannitol. The blastomeres were fused following a short electric pulse at 1.2 kV/cm and 4µs using Electro cell manipulator BTX (ECM 2001). After the electric pulse, the fused embryos were scored and further cultured in 50 µl drops of RVCL media supplemented with 1% BSA for 48 hr in humidified atmosphere of 5% CO2 at 38.5ºC in CO2 incubator. The morulae were used for ESCs microinjections.

Microinjection of ES cells into tetraploid ivf embryos

The procedure was carried out as per Kharche et al. [10], with modifications on the stage of an inverted microscope (NIKON, Eclipse, TE 2000U) at 40x, 100x, and 200x magnification, using Hoffman Modulation Contrast (HMC) optics. The microscope had a camera (DXM 1200, Nikon Japan) that allowed the procedure to be followed on a computer monitor and for taking pictures to the computer. The microscope was equipped with three axis hanging joystick oil Hydraulic Micromanipulator (NT-88-V3, Narishige, Japan) and with microinjector (IM-9B, Narishige, Japan) and pneumatic injector (IM-9C, Narishige, Japan). The above micromanipulator was equipped for piezoxpert (Eppendorf AG 22331 Hamburg Germany) for smoother injection so it caused minimal damage to the zonapellucida of the embryo for optimum survival of embryos. Coarse adjustment was done at 40x magnification followed by fine adjustment at 200x magnification. 60 mm culture dish was used as a microinjection chamber. Two rows consisting of 10 µL droplets for IVF tetraploid morulae were placed right to the centre of the dish and two rows consisting of 10 µL droplets for embryonic stem cell colonies were placed on the left to the centre of the dish. These droplets were covered with mineral oil.

Briefly, an ESC like cell colony was picked up, into the injection pipette. This colony was pushed forward until it was near the tip of the pipette, the pipette was advanced quickly towards the droplet containing morula where it was held with holding pipette. The injection pipette containing ESC at the very tip of the pipette was injected in to the zonapellucida by applying piezo electric pulse.The micropipette was then pushed through the zonapellucida. When the pipette was well within the embryo, a gentle suction was applied followed by a sudden rush of cytoplasm into the pipette as the plasma membrane ruptured. Some cytoplasm was seen in the pipette and the ESC moved backwards into the pipette. The ESC and the cytoplasm were pushed back into the morula so that the ESC lies within the morula and pipette was then retracted from the morula. The injected morula was then released from the holding pipette. We routinely applied a light negative pressure to the pipette during its withdrawal. This appeared to increase the survival rate of the embryos [11]. The injected embryos were further cultured in 50 µl drops of RVCL media supplemented with 1% BSA for 48 hr in humidified atmosphere of 5% CO2 at 38.5?C in CO2 incubator.

Embryo transfer

In vitro produced embryos were transferred in two non descript Sirohi goats at naturaloestrus. Recipient was deprived of feed and water for 24 hand put under general anesthesia using xylazine (0.2 mg/kgbody weight) and ketamine (4.4 - 6.6 mg/kg body weight).The reproductive tract was exteriorized through a mid-ventralincision to allow visual confirmation of a corpus luteum/lutea on ovaries. Chimeric embryos were transferred surgically at the tip of uterine horn ipsilateral to the ovary containing corpus luteum of two naturally synchronized surrogate doe (4 embryos each).


Parthenogenetic activation and embryonic stem cell

The percentage of 2 cell, 4 cell, 8-16 cell, morula, hatched blastocyst and cleavage rate of parthenogenetic goat embryos were 20.03 ± 5.09%, 20.02 ± 6.30%, 24.41 ± 4.12%, 25.61 ± 7.63%, 9.92 ± 3.97% and 72.78 ± 13.39%, respectively. Furthermore, 75.55 ± 12.37% embryonic stem cell colonies were formed from hatched blastocysts.

In vitro fertilization and tetraploid embryos

The cleavage rate following IVF was 37.08 ± 4.34%. Furthermore, electrofusion of 2 cell embryo resulted in 75.09 ± 7.86% fusion rate and 86.5 ± 13.02% cleavage rate. The percentage of 2 cell, 4 cell, 8-16 cell and morula production of tetraploid embryos were 00 ± 00%, 11.33 ± 7.85%, 50.66 ± 14.54% and 38 ± 11.3%, respectively.

Microinjection of ES cells into tetraploid ivf embryos and embryo transfers

In this study 8 microinjected chimeric embryos were transferred into two goats as per the method of Kharche et al. [8]. Following transfer, none of the recipients were found pregnant at day 35th post transfer by ultrasonography. Tetraploid (4n) embryos are used to produce chimeras with embryonic stem cells (ESCs) and diploid (2n) embryos, which have been used to substitute microinjection of DNA and nuclear transfer for the creation of genetically modified animals [12,13].

Although tetraploid embryos can form blastocysts, their post-implantation development is impaired because of the absence of epiblast cells and the failure of embryos to survive beyond mid-gestation [14]. Tetraploid and diploid embryos can be aggregated/hybridized to make chimeras. Within this construction, tetraploid cells rarely contribute to the embryo itself (which is derived from the epiblast); rather, they contribute mainly to the hypoblast and the trophectoderm [15,16]. Therefore, chimerism of parthenogenetic ES cells and tetraploid embryos can been used in the tetraploid complementation assay (TCA) for production of parthenogenetic goat.

In mouse, ES-tetraploid chimeras were reported by Nagy et al. [4], and according to him the newborns were almost all completely ES-derived, as judged by CPI isozyme analysis, but tetraploid cells were found in the yolk sac endoderm and trophectoderm lineage. Although newborns failed to survive after birth, despite they had normal birthweight and anatomically they appeared normal. In domestic animals, chimeric calves from ES-like cells aggregated with tetraploid embryos were reported to have been born [17,18], however, ES like cells contributed to chimera formation only at a very low level.


In conclusion, the results presented here indicate that caprine ES-like cells can be incorporated into IVF tetraploid embryos by microinjection so as to produce chimeric embryos. However, in vivo development of chimeric embryos was not achieved.


The authors wish to thanks to the ADG, NFBSFARA, New Delhi for providing the funding and Director, C.I.R.G., Makhdoom, Farah, Mathura, UP, India for providing the facilities needed.


1. Srirattana K, Sripunya N, Sangmalee A, Imsoonthornruksa S, Liang Y, Ketudat-Cairns M, et al. Developmental potential of vitrified goat oocytes following somatic cell nuclear transfer and parthenogenetic activation. Small Ruminant Research. 2013; 112: 141-146.

2. Pathak J, Kharche SD, Goel AK, Jindal SK. A comparative study on parthenogenetic activation and embryo production from in vitro matured caprine oocytes. Small Ruminant Research. 2013; 113: 136- 140.

3. De LFR, King WA. Developmental consequences of karyokinesis without cytokinesis during the first mitotic cell cycle of bovine parthenotes. Biol Reprod. 1998; 58: 952-962.

4. Nagy A, Gocza E, Diaz EM, Prideaux VR, Ivanyi E, Markkula M, et al. Embryonic stem cells alone are able to support fetal development in the mouse. Development. 1990; 110: 815-821.

5. Nagy A, Rossant J, Nagy R, Abramow-Newerly W, Roder JC. Derivation of completely cell culture-derived mice from earlypassage embryonic stem cells. Proc Natl Acad Sci USA. 1993; 90: 8424-8428.

6. Ueda O, Jishage K, Kamada N, Uchida S, Suzuki H. Production of mice entirely derived from embryonic stem (ES) cell with many passages by coculture of ES cells with cytochalasin B induced tetraploid embryos. Exp Anim. 1995; 44: 205-210.

7. Kharche SD, Goel AK, Jindal SK, Sinha NK, Yadav P. Effect of somatic cells co-cultures on cleavage and development of in vitro fertilized caprine embryos. Indian J Anim Sci. 2008; 78: 686-692.

8. Kharche SD, Goel AK, Jindal SK, Goel P, Jha BK. Birth of twin kids following transfer of in-vitro produced caprine embryos. Indian J Anim Sci. 2011; 81: 1132-1134.

9. Parrish JJ, Susko-Parrish JL, Leibfned-Rutledge ML, Critser ES, Eyestone WH, First NL. Bovine in vitro fertilization with frozenthawed semen.Theriogenol.1986; 25: 591-600.

10. Kharche S, Pathak J, Agarwal S, Kushwah B, Sikarwar AKS. Effect of CaIonophoreOn Blastocyst Production Following Intracytoplasmic Sperm Injection in Caprine Oocytes. Reprod Dom Anim. 2016; 51: 611-617.

11. Kimura Y, Yanagimachi R. Intracytoplasmic sperm injection in the mouse. Biol Reprod. 1995; 52: 709-720.

12. Wang ZQ, Kiefer F, Urbanek P, Wagner EF. Generation of completely embryonic stem cell-derived mutant mice using tetraploid blastocyst injection. Mech Dev. 1997; 62: 137-145.

13. Mueller S, Prelle K, Rieger N, Petznek H, Lassnig C, Luksch U, et al. Chimeric pigs following blastocyst injection of transgenic porcine primordial germ cells. Mol Reprod Dev. 1999; 54: 244-254.

14. Kaufman MH, Webb S. Postimplantation development of tetraploid mouse embryos produced by electrofusion. Development. 1990; 110: 1121-1132.

15. Tarkowski AK, Witkowska A, Opas J. Development of cytochalasin B-induced tetraploid and diploid/tetraploid mosaic mouse embryos. J Embryol Exp Morphol. 1977; 41: 47-64.

16. James RM, Klerkx AH, Keighren M, Flockhart JH, West JD. Restricted distribution of tetraploid cells in mouse tetraploid-diploid chimaeras. Dev Biol. 1995; 167: 213-226.

17. Iwasaki S, Campbell KH, Galli C, Akiyama K. Production of live calves derived from embryonic stem-like cells aggregated with tetraploid embryos. Biol Reprod. 2000; 62: 470-475.

18. Saito S, Sawai K, Ugai H, Moriyasu S, Minamihashi A, Yamamoto Y, et al. Generation of cloned calves and transgenic chimeric embryos from bovine embryonic stem-like cells. Biochemical and Biophysical Research Communications. 2003; 309: 104-113.

Kharche SD, Pathak J, Sikarwar AS, Gangwar C, Priyadharsini R, et al. (2017) In vivo Transfer of Reconstructed Goat Chimeric Embryos Produced by Microinjection of Embryonic Stem Cells into Tetraploid IVF Embryos. JSM Cell Dev Biol 5(1): 1021.

Received : 04 Apr 2017
Accepted : 25 May 2017
Published : 28 May 2017
Annals of Otolaryngology and Rhinology
ISSN : 2379-948X
Launched : 2014
JSM Schizophrenia
Launched : 2016
Journal of Nausea
Launched : 2020
JSM Internal Medicine
Launched : 2016
JSM Hepatitis
Launched : 2016
JSM Oro Facial Surgeries
ISSN : 2578-3211
Launched : 2016
Journal of Human Nutrition and Food Science
ISSN : 2333-6706
Launched : 2013
JSM Regenerative Medicine and Bioengineering
ISSN : 2379-0490
Launched : 2013
JSM Spine
ISSN : 2578-3181
Launched : 2016
Archives of Palliative Care
ISSN : 2573-1165
Launched : 2016
JSM Nutritional Disorders
ISSN : 2578-3203
Launched : 2017
Annals of Neurodegenerative Disorders
ISSN : 2476-2032
Launched : 2016
Journal of Fever
ISSN : 2641-7782
Launched : 2017
JSM Bone Marrow Research
ISSN : 2578-3351
Launched : 2016
JSM Mathematics and Statistics
ISSN : 2578-3173
Launched : 2014
Journal of Autoimmunity and Research
ISSN : 2573-1173
Launched : 2014
JSM Arthritis
ISSN : 2475-9155
Launched : 2016
JSM Head and Neck Cancer-Cases and Reviews
ISSN : 2573-1610
Launched : 2016
JSM General Surgery Cases and Images
ISSN : 2573-1564
Launched : 2016
JSM Anatomy and Physiology
ISSN : 2573-1262
Launched : 2016
JSM Dental Surgery
ISSN : 2573-1548
Launched : 2016
Annals of Emergency Surgery
ISSN : 2573-1017
Launched : 2016
Annals of Mens Health and Wellness
ISSN : 2641-7707
Launched : 2017
Journal of Preventive Medicine and Health Care
ISSN : 2576-0084
Launched : 2018
Journal of Chronic Diseases and Management
ISSN : 2573-1300
Launched : 2016
Annals of Vaccines and Immunization
ISSN : 2378-9379
Launched : 2014
JSM Heart Surgery Cases and Images
ISSN : 2578-3157
Launched : 2016
Annals of Reproductive Medicine and Treatment
ISSN : 2573-1092
Launched : 2016
JSM Brain Science
ISSN : 2573-1289
Launched : 2016
JSM Biomarkers
ISSN : 2578-3815
Launched : 2014
JSM Biology
ISSN : 2475-9392
Launched : 2016
Archives of Stem Cell and Research
ISSN : 2578-3580
Launched : 2014
Annals of Clinical and Medical Microbiology
ISSN : 2578-3629
Launched : 2014
JSM Pediatric Surgery
ISSN : 2578-3149
Launched : 2017
Journal of Memory Disorder and Rehabilitation
ISSN : 2578-319X
Launched : 2016
JSM Tropical Medicine and Research
ISSN : 2578-3165
Launched : 2016
JSM Head and Face Medicine
ISSN : 2578-3793
Launched : 2016
JSM Cardiothoracic Surgery
ISSN : 2573-1297
Launched : 2016
JSM Bone and Joint Diseases
ISSN : 2578-3351
Launched : 2017
JSM Bioavailability and Bioequivalence
ISSN : 2641-7812
Launched : 2017
JSM Atherosclerosis
ISSN : 2573-1270
Launched : 2016
Journal of Genitourinary Disorders
ISSN : 2641-7790
Launched : 2017
Journal of Fractures and Sprains
ISSN : 2578-3831
Launched : 2016
Journal of Autism and Epilepsy
ISSN : 2641-7774
Launched : 2016
Annals of Marine Biology and Research
ISSN : 2573-105X
Launched : 2014
JSM Health Education & Primary Health Care
ISSN : 2578-3777
Launched : 2016
JSM Communication Disorders
ISSN : 2578-3807
Launched : 2016
Annals of Musculoskeletal Disorders
ISSN : 2578-3599
Launched : 2016
Annals of Virology and Research
ISSN : 2573-1122
Launched : 2014
JSM Renal Medicine
ISSN : 2573-1637
Launched : 2016
Journal of Muscle Health
ISSN : 2578-3823
Launched : 2016
JSM Genetics and Genomics
ISSN : 2334-1823
Launched : 2013
JSM Anxiety and Depression
ISSN : 2475-9139
Launched : 2016
Clinical Journal of Heart Diseases
ISSN : 2641-7766
Launched : 2016
Annals of Medicinal Chemistry and Research
ISSN : 2378-9336
Launched : 2014
JSM Pain and Management
ISSN : 2578-3378
Launched : 2016
JSM Women's Health
ISSN : 2578-3696
Launched : 2016
Clinical Research in HIV or AIDS
ISSN : 2374-0094
Launched : 2013
Journal of Endocrinology, Diabetes and Obesity
ISSN : 2333-6692
Launched : 2013
Journal of Substance Abuse and Alcoholism
ISSN : 2373-9363
Launched : 2013
JSM Neurosurgery and Spine
ISSN : 2373-9479
Launched : 2013
Journal of Liver and Clinical Research
ISSN : 2379-0830
Launched : 2014
Journal of Drug Design and Research
ISSN : 2379-089X
Launched : 2014
JSM Clinical Oncology and Research
ISSN : 2373-938X
Launched : 2013
JSM Bioinformatics, Genomics and Proteomics
ISSN : 2576-1102
Launched : 2014
JSM Chemistry
ISSN : 2334-1831
Launched : 2013
Journal of Trauma and Care
ISSN : 2573-1246
Launched : 2014
JSM Surgical Oncology and Research
ISSN : 2578-3688
Launched : 2016
Annals of Food Processing and Preservation
ISSN : 2573-1033
Launched : 2016
Journal of Radiology and Radiation Therapy
ISSN : 2333-7095
Launched : 2013
JSM Physical Medicine and Rehabilitation
ISSN : 2578-3572
Launched : 2016
Annals of Clinical Pathology
ISSN : 2373-9282
Launched : 2013
Annals of Cardiovascular Diseases
ISSN : 2641-7731
Launched : 2016
Journal of Behavior
ISSN : 2576-0076
Launched : 2016
Annals of Clinical and Experimental Metabolism
ISSN : 2572-2492
Launched : 2016
Clinical Research in Infectious Diseases
ISSN : 2379-0636
Launched : 2013
JSM Microbiology
ISSN : 2333-6455
Launched : 2013
Journal of Urology and Research
ISSN : 2379-951X
Launched : 2014
Journal of Family Medicine and Community Health
ISSN : 2379-0547
Launched : 2013
Annals of Pregnancy and Care
ISSN : 2578-336X
Launched : 2017
Annals of Aquaculture and Research
ISSN : 2379-0881
Launched : 2014
Clinical Research in Pulmonology
ISSN : 2333-6625
Launched : 2013
Journal of Immunology and Clinical Research
ISSN : 2333-6714
Launched : 2013
Annals of Forensic Research and Analysis
ISSN : 2378-9476
Launched : 2014
JSM Biochemistry and Molecular Biology
ISSN : 2333-7109
Launched : 2013
Annals of Breast Cancer Research
ISSN : 2641-7685
Launched : 2016
Annals of Gerontology and Geriatric Research
ISSN : 2378-9409
Launched : 2014
Journal of Sleep Medicine and Disorders
ISSN : 2379-0822
Launched : 2014
JSM Burns and Trauma
ISSN : 2475-9406
Launched : 2016
Chemical Engineering and Process Techniques
ISSN : 2333-6633
Launched : 2013
Annals of Clinical Cytology and Pathology
ISSN : 2475-9430
Launched : 2014
JSM Allergy and Asthma
ISSN : 2573-1254
Launched : 2016
Journal of Neurological Disorders and Stroke
ISSN : 2334-2307
Launched : 2013
Annals of Sports Medicine and Research
ISSN : 2379-0571
Launched : 2014
JSM Sexual Medicine
ISSN : 2578-3718
Launched : 2016
Annals of Vascular Medicine and Research
ISSN : 2378-9344
Launched : 2014
JSM Biotechnology and Biomedical Engineering
ISSN : 2333-7117
Launched : 2013
Journal of Hematology and Transfusion
ISSN : 2333-6684
Launched : 2013
JSM Environmental Science and Ecology
ISSN : 2333-7141
Launched : 2013
Journal of Cardiology and Clinical Research
ISSN : 2333-6676
Launched : 2013
JSM Nanotechnology and Nanomedicine
ISSN : 2334-1815
Launched : 2013
Journal of Ear, Nose and Throat Disorders
ISSN : 2475-9473
Launched : 2016
JSM Ophthalmology
ISSN : 2333-6447
Launched : 2013
Journal of Pharmacology and Clinical Toxicology
ISSN : 2333-7079
Launched : 2013
Annals of Psychiatry and Mental Health
ISSN : 2374-0124
Launched : 2013
Medical Journal of Obstetrics and Gynecology
ISSN : 2333-6439
Launched : 2013
Annals of Pediatrics and Child Health
ISSN : 2373-9312
Launched : 2013
JSM Clinical Pharmaceutics
ISSN : 2379-9498
Launched : 2014
JSM Foot and Ankle
ISSN : 2475-9112
Launched : 2016
JSM Alzheimer's Disease and Related Dementia
ISSN : 2378-9565
Launched : 2014
Journal of Addiction Medicine and Therapy
ISSN : 2333-665X
Launched : 2013
Journal of Veterinary Medicine and Research
ISSN : 2378-931X
Launched : 2013
Annals of Public Health and Research
ISSN : 2378-9328
Launched : 2014
Annals of Orthopedics and Rheumatology
ISSN : 2373-9290
Launched : 2013
Journal of Clinical Nephrology and Research
ISSN : 2379-0652
Launched : 2014
Annals of Community Medicine and Practice
ISSN : 2475-9465
Launched : 2014
Annals of Biometrics and Biostatistics
ISSN : 2374-0116
Launched : 2013
JSM Clinical Case Reports
ISSN : 2373-9819
Launched : 2013
Journal of Cancer Biology and Research
ISSN : 2373-9436
Launched : 2013
Journal of Surgery and Transplantation Science
ISSN : 2379-0911
Launched : 2013
Journal of Dermatology and Clinical Research
ISSN : 2373-9371
Launched : 2013
JSM Gastroenterology and Hepatology
ISSN : 2373-9487
Launched : 2013
TEST Journal of Dentistry
ISSN : 1234-5678
Launched : 2014
Annals of Nursing and Practice
ISSN : 2379-9501
Launched : 2014
JSM Dentistry
ISSN : 2333-7133
Launched : 2013
Author Information X