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  1. #1
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    Default In vitro culturing and harvesting of human plucked hair follicles

    http://corescholar.libraries.wright....69&context=jbm



    ABSTRACT
    Human hair follicles are miniature hair growing organs. A common human hair disorder
    is androgenetic alopecia (AGA), which becomes a medical problem only when the hair loss is
    subjectively seen as excessive, premature, and distressing on the scalp. The objectives of the
    present study were to culture the hair stem cells in vitro and to study the morphology of the
    cultured cells for the treatment of AGA. The present study proposes that plucked human hairs are
    a cheap source to treat male baldness and in vitro culturing of hair follicle cells is the potential
    method to apply the cultured cells back into the balding scalp. It may be possible to create
    thousands of hair follicles from that original follicle. In this study human hair follicle cells of normal and AGA male groups were taken by plucking the hair follicle cells. The hair follicles cells of normal and AGA were cultured in vitro without a feeder layer, as the feeder layer has many drawbacks. The plucked hair follicles which were in the anagen stage were selected from both groups. These hair follicles were digested by enzymatic disaggregation using trypsin/EDTA. Then these cells were cultured in a FAD medium ( Dulbecco’s Modified Eagle Medium and Ham’s F 12 medium, 3:1) plus a 17% serum and incubated in a CO2 Incubator at 37 °C in 5% CO2 without a feeder layer. The whole procedure was performed under sterile conditions. The morphology of cultured and subcultured cells was observed daily under a phase contrast microscope for 14 days. The viable cultured cells of both groups refracted light, while dead cells appeared black. Keratinocytes appeared after 24 hours, Melanocytes appeared at 48 hours, and stem cells appeared in 7 to 10 days. Shelf life of cultured and subcultured cells of normal and AGA group was 12 and 7 days, respectively. Live cell counting was done by using an improved Neubauer chamber. DNA extraction and optical density (OD) assay of cultured and subcultured cells was undertaken using the plucked hair follicles of normal and AGA human male subject. The plucked human hair follicles cells were harvested and cultured successfully without a feeder layer. Their genomic DNA was extracted successfully. This hair cloning technique is an alternative to the usual method of hair transplantation. The most positive aspect of the new technique compared to hair transplantation is the preservation of the 'donor hair area'. This technique will be cheaper and more 'patient friendly'.

  2. #2
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    Not sure why this has got no replies yet given the statement:

    "This hair cloning technique is an alternative to the usual method of hair transplantation. The most positive aspect of the new technique compared to hair transplantation is the preservation of the 'donor hair area'. This technique will be cheaper and more 'patient friendly'."

    Is there something implausible about this?

  3. #3
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    Wow, interesting, the protocol is cheap, safe, and the "cells were harvested and cultured successfully without a feeder layer" and "Their genomic DNA was extracted successfully"

    "The viable cultured cells of both groups refracted light, while dead cells appeared black. Keratinocytes appeared after 24 hours, Melanocytes appeared at 48 hours, and stem cells appeared in 7 to 10 days"

    We need them to inject those cells see if it can grow a new hair. If yes, they are right it's 'hair cloning' as they claim. But we would need confirmation of that with them putting back those cells in a patient.

    the 3 researchers are from a big Pakistan's university, maybe they are trying/will try soon to have the confirmation

    But even if it can't grow a new hair, it could still be a helping protocol, as these cells would at least help the current hair and the miniaturized ones

    we would need more info from them

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    Will have a look to find out if I can find contact info. I didn't see this posted tbf.

  5. #5
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    i don't find contact mail too

    Iram Rehman, Quaid-i-Azam University, Islamabad, Pakistan
    Attika Rehman, Quaid-i-Azam University, Islamabad, Pakistan
    Samina Jalali, Quaid-i-Azam University, Islamabad, Pakistan

    But I see that Attika Rehman is a MPhil student from department of biochemistry ( i don't see the two others in the staff or students so they are working in another department of the Univ)

    http://bch.qau.edu.pk/enrollstdmp.php

    And there's a mail of the departement : bch@qau.edu.pk (and phone number), maybe they can give us the personal mail of one of the three involved


    Regarding the Univ: ( from wikipedia)

    "The Quaid-i-Azam University, (formerly known as Islamabad University) is a public research university located in Islamabad, Pakistan.[3] Consisting of four faculties, nine affiliated research institutes, 38 departments, the university is consistently ranked as the top university in Pakistan "[4] by the HEC and is ranked by Quacquarelli Symonds ranking among the top 500 universities in the world in 2013, while it was ranked at 69th top university of the world in field of 'Engineering and Technology'"


    So they are def not looking like jokers, but we need them to inject those cells on a scalp for confirmation of their high claims (maybe only predictions). But the protocol is safe and it's take place in pakistan (also near india), they should have the permission to do so as it's like other basic cellular therapy

  6. #6
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    what? they are just plucking hairs and culturing those cells or what?
    we're not talking about DP or DSC cells here, right?

    i thought it was already proven that only those two cell types can induce new follicles. are they now trying to just culture simple hair fibre cells?

  7. #7
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    Yes Joachim

    but apparently with their protocol and way of culturing;

    "Keratinocytes appeared after 24 hours, Melanocytes appeared at 48 hours, and stem cells appeared in 7 to 10 days"

    but what are these stem cells they talk about? DS? DP? or maybe HF dermal stem cells ?

    "Hair follicle dermal stem cells regenerate the dermal sheath, repopulate the dermal papilla, and modulate hair type"

    http://www.ncbi.nlm.nih.gov/pubmed/25465495


    The dermal papilla (DP) provide instructive signals required to activate epithelial progenitors and initiate hair follicle regeneration. DP cell numbers fluctuate over the hair cycle, and hair loss is associated with gradual depletion/atrophy of DP cells. How DP cell numbers are maintained in healthy follicles remains unclear. We performed in vivo fate mapping of adult hair follicle dermal sheath (DS) cells to determine their lineage relationship with DP and found that a subset of DS cells are retained following each hair cycle, exhibit self-renewal, and repopulate the DS and the DP with new cells. Ablating these hair follicle dermal stem cells and their progeny retarded hair regrowth and altered hair type specification, suggesting that they function to modulate normal DP function. This work identifies a bipotent stem cell within the adult hair follicle mesenchyme and has important implications toward restoration of hair growth after injury, disease, and aging."



    What are the stem cells they are seeing at 7 days because of their protocol? it's the question, if those are powerful cells, as they seem to claim, then their plucked protocol is insanely interesting

  8. #8
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    Quote Originally Posted by lacazette View Post
    Yes Joachim

    but apparently with their protocol and way of culturing;

    "Keratinocytes appeared after 24 hours, Melanocytes appeared at 48 hours, and stem cells appeared in 7 to 10 days"

    but what are these stem cells they talk about? DS? DP? or maybe HF dermal stem cells ?

    "Hair follicle dermal stem cells regenerate the dermal sheath, repopulate the dermal papilla, and modulate hair type"

    http://www.ncbi.nlm.nih.gov/pubmed/25465495


    The dermal papilla (DP) provide instructive signals required to activate epithelial progenitors and initiate hair follicle regeneration. DP cell numbers fluctuate over the hair cycle, and hair loss is associated with gradual depletion/atrophy of DP cells. How DP cell numbers are maintained in healthy follicles remains unclear. We performed in vivo fate mapping of adult hair follicle dermal sheath (DS) cells to determine their lineage relationship with DP and found that a subset of DS cells are retained following each hair cycle, exhibit self-renewal, and repopulate the DS and the DP with new cells. Ablating these hair follicle dermal stem cells and their progeny retarded hair regrowth and altered hair type specification, suggesting that they function to modulate normal DP function. This work identifies a bipotent stem cell within the adult hair follicle mesenchyme and has important implications toward restoration of hair growth after injury, disease, and aging."



    What are the stem cells they are seeing at 7 days because of their protocol? it's the question, if those are powerful cells, as they seem to claim, then their plucked protocol is insanely interesting

    It's strange they didn't implant them back in a human head though...

    It's common for researchers to break up their research into multiple papers though (boosts the appearance of their productivity), so maybe they have a follow up paper coming out shortly (hopefully).

    One would imagine if they came up with the holy grail, they might want to capitalize off of it sooner than later.

    Also the work from this paper could have happened upwards of six months ago, so who knows where they are now.....

    Maybe some googling of their names for news/company affiliations... or uni blog... or reaching out and contacting them

  9. #9
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    Sounds similar to histogen... hoping a cure comes out before fin starts losing efficiency lol

  10. #10
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    We should convince Dr Gho to try this protocol as he desperatly try hair cloning for more than a decade, he could try serious stuff now ^^

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