PMID- 35249016
OWN - NLM
STAT- MEDLINE
DCOM- 20230605
LR  - 20230609
IS  - 1422-6421 (Electronic)
IS  - 1422-6405 (Linking)
VI  - 212
IP  - 3
DP  - 2023
TI  - Successful in vivo Transplantation of Cultured and Enriched Testicular Germ Cells 
      of Pre-Pubertal Bucks to Busulfan-Treated Homologous Recipients.
PG  - 232-244
LID - 10.1159/000523891 [doi]
AB  - The objective of the present study was to establish a workable approach for the 
      production of germ cell (GC)-depleted recipient goat model using intra-testicular 
      busulfan treatment and transplantation of cultured and enriched caprine-male GC 
      (cmGCs) into the homologous recipients under ultrasonography (USG) guidance. The 
      evaluation of post-transplantation colonization of donor cmGCs and restoration of 
      the normal architecture of seminiferous tubules (ST) was performed. For this, the 
      cmGCs of pre-pubertal male goats were isolated and enriched by differential 
      platting for culture until the third passage. Thereafter, cells were harvested 
      and further enriched by magnetic-activated cell sorting using rabbit-anti-CD90 
      antibody. After confirmation of metabolic viability (MTT-assay) and 
      cluster-forming ability (crystal violet staining) of CD90+ cmGCs, the cells were 
      labeled with a lipophilic red-fluorescent dye (PKH26) before transplanted into 
      the recipient male goats by injection directly into the mediastinum testes under 
      USG guidance. The colonization and repopulation of transplanted CD90+ cmGCs into 
      the recipient ST was observed up to 8 weeks post-transplantation. The 
      PKH26-labeled donor cell-derived colonies were identified in enzymatically 
      digested ST and cryosections of recipient testes. Moreover, histochemical 
      analyses revealed the restoration of the normal architecture of ST of recipient 
      testis after GC transplantation. Therefore, the results suggest that the 
      reproductive competence of infertile animals can be restored through mGC therapy 
      and thus the methodology presented herein could be useful to obtain donor 
      mGCs-derived functional male gametes in the recipient animal testis.
CI  - (c) 2022 S. Karger AG, Basel.
FAU - Singh, Shiva P
AU  - Singh SP
AD  - Animal Physiology and Reproduction Division, ICAR-Central Institute for Research 
      on Goats, Mathura, India.
FAU - Kharche, Suresh D
AU  - Kharche SD
AD  - Animal Physiology and Reproduction Division, ICAR-Central Institute for Research 
      on Goats, Mathura, India.
FAU - Soni, Yogesh K
AU  - Soni YK
AD  - Animal Physiology and Reproduction Division, ICAR-Central Institute for Research 
      on Goats, Mathura, India.
FAU - Pathak, Manisha
AU  - Pathak M
AD  - Animal Physiology and Reproduction Division, ICAR-Central Institute for Research 
      on Goats, Mathura, India.
FAU - Ranjan, Ravi
AU  - Ranjan R
AD  - Animal Physiology and Reproduction Division, ICAR-Central Institute for Research 
      on Goats, Mathura, India.
FAU - Majhi, Sullip K
AU  - Majhi SK
AD  - ICAR-National Bureau of Fish Genetic Resources, Lucknow, India.
FAU - Pawaiya, Rajveer S
AU  - Pawaiya RS
AD  - Animal Health Division, ICAR-Central Institute for Research on Goats, Mathura, 
      India.
FAU - Singh, Manoj K
AU  - Singh MK
AD  - Animal Genetics and Breeding Division, ICAR-Central Institute for Research on 
      Goats, Mathura, India.
FAU - Chauhan, Manmohan S
AU  - Chauhan MS
AD  - ICAR-National Dairy Research Institute, Karnal, India.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220304
PL  - Switzerland
TA  - Cells Tissues Organs
JT  - Cells, tissues, organs
JID - 100883360
RN  - 0 (PKH 26)
RN  - G1LN9045DK (Busulfan)
SB  - IM
MH  - Animals
MH  - Male
MH  - Rabbits
MH  - *Testis
MH  - *Busulfan/pharmacology
MH  - Spermatogenesis
MH  - Goats
MH  - Germ Cells
MH  - Spermatogonia
OTO - NOTNLM
OT  - Busulfan
OT  - Germ cells
OT  - Goats
OT  - Homologous transplantation
OT  - Intra-testicular injection
EDAT- 2022/03/07 06:00
MHDA- 2023/06/05 06:42
CRDT- 2022/03/06 20:37
PHST- 2022/01/12 00:00 [received]
PHST- 2022/02/27 00:00 [accepted]
PHST- 2023/06/05 06:42 [medline]
PHST- 2022/03/07 06:00 [pubmed]
PHST- 2022/03/06 20:37 [entrez]
AID - 000523891 [pii]
AID - 10.1159/000523891 [doi]
PST - ppublish
SO  - Cells Tissues Organs. 2023;212(3):232-244. doi: 10.1159/000523891. Epub 2022 Mar 
      4.